Biolistics: Techniques where DNA is delivered directly into organelles at high speed using projectiles coated with nucleic acid, shot from a helium-powered gun (gene gun). One of these techniques involves immunization by DNA VACCINES, which delivers DNA-coated gold beads to the epidermis.Helium: Helium. A noble gas with the atomic symbol He, atomic number 2, and atomic weight 4.003. It is a colorless, odorless, tasteless gas that is not combustible and does not support combustion. It was first detected in the sun and is now obtained from natural gas. Medically it is used as a diluent for other gases, being especially useful with oxygen in the treatment of certain cases of respiratory obstruction, and as a vehicle for general anesthetics. (Dorland, 27th ed)Chickens: Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.Pressure: A type of stress exerted uniformly in all directions. Its measure is the force exerted per unit area. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)Brachiaria: A plant genus of the family POACEAE originating from the savanna of eastern Africa. It is widely grown for livestock forage.Vacuum: A space in which the pressure is far below atmospheric pressure so that the remaining gases do not affect processes being carried on in the space.Chick Embryo: The developmental entity of a fertilized chicken egg (ZYGOTE). The developmental process begins about 24 h before the egg is laid at the BLASTODISC, a small whitish spot on the surface of the EGG YOLK. After 21 days of incubation, the embryo is fully developed before hatching.Plants, Genetically Modified: PLANTS, or their progeny, whose GENOME has been altered by GENETIC ENGINEERING.RNA Interference: A gene silencing phenomenon whereby specific dsRNAs (RNA, DOUBLE-STRANDED) trigger the degradation of homologous mRNA (RNA, MESSENGER). The specific dsRNAs are processed into SMALL INTERFERING RNA (siRNA) which serves as a guide for cleavage of the homologous mRNA in the RNA-INDUCED SILENCING COMPLEX. DNA METHYLATION may also be triggered during this process.Plant Leaves: Expanded structures, usually green, of vascular plants, characteristically consisting of a bladelike expansion attached to a stem, and functioning as the principal organ of photosynthesis and transpiration. (American Heritage Dictionary, 2d ed)Plant Proteins: Proteins found in plants (flowers, herbs, shrubs, trees, etc.). The concept does not include proteins found in vegetables for which VEGETABLE PROTEINS is available.Plant Roots: The usually underground portions of a plant that serve as support, store food, and through which water and mineral nutrients enter the plant. (From American Heritage Dictionary, 1982; Concise Dictionary of Biology, 1990)Genes, Plant: The functional hereditary units of PLANTS.Plant Extracts: Concentrated pharmaceutical preparations of plants obtained by removing active constituents with a suitable solvent, which is evaporated away, and adjusting the residue to a prescribed standard.Laboratory ManualsManuals as Topic: Books designed to give factual information or instructions.Periodicals as Topic: A publication issued at stated, more or less regular, intervals.HandbooksPublishing: "The business or profession of the commercial production and issuance of literature" (Webster's 3d). It includes the publisher, publication processes, editing and editors. Production may be by conventional printing methods or by electronic publishing.Histology: The study of the structure of various TISSUES of organisms on a microscopic level.Andrology: A scientific or medical discipline concerning the study of male reproductive biology, diseases of the male genital organs, and male infertility. Major areas of interest include ENDOCRINOLOGY; SPERMATOGENESIS; semen analysis; FERTILIZATION; CONTRACEPTION; and CRYOPRESERVATION.Soybeans: An annual legume. The SEEDS of this plant are edible and used to produce a variety of SOY FOODS.Fertility: The capacity to conceive or to induce conception. It may refer to either the male or female.Ovule: The element in plants that contains the female GAMETOPHYTES.Plant Infertility: The failure of PLANTS to complete fertilization and obtain seed (SEEDS) as a result of defective POLLEN or ovules, or other aberrations. (Dict. of Plant Genet. and Mol. Biol., 1998)Pollen: The fertilizing element of plants that contains the male GAMETOPHYTES.Germ Cells, Plant: The reproductive cells of plants.Oleic Acid: An unsaturated fatty acid that is the most widely distributed and abundant fatty acid in nature. It is used commercially in the preparation of oleates and lotions, and as a pharmaceutical solvent. (Stedman, 26th ed)Oleic Acids: A group of fatty acids that contain 18 carbon atoms and a double bond at the omega 9 carbon.Glycine: A non-essential amino acid. It is found primarily in gelatin and silk fibroin and used therapeutically as a nutrient. It is also a fast inhibitory neurotransmitter.Plant Oils: Oils derived from plants or plant products.Apiaceae: A large plant family in the order Apiales, also known as Umbelliferae. Most are aromatic herbs with alternate, feather-divided leaves that are sheathed at the base. The flowers often form a conspicuous flat-topped umbel. Each small individual flower is usually bisexual, with five sepals, five petals, and an enlarged disk at the base of the style. The fruits are ridged and are composed of two parts that split open at maturity.Potyviridae: A family of RNA plant viruses with flexuous, filamentous particles and consisting of six genera: POTYVIRUS; Ipomovirus; Macluravirus; Rymovirus; Tritimovirus; and Bymovirus. All members of the family form cytoplasmic cylindrical inclusion bodies during infection.Triticum: A plant genus of the family POACEAE that is the source of EDIBLE GRAIN. A hybrid with rye (SECALE CEREALE) is called TRITICALE. The seed is ground into FLOUR and used to make BREAD, and is the source of WHEAT GERM AGGLUTININS.Mosaic Viruses: Viruses which produce a mottled appearance of the leaves of plants.Plant Diseases: Diseases of plants.MicroRNAs: Small double-stranded, non-protein coding RNAs, 21-25 nucleotides in length generated from single-stranded microRNA gene transcripts by the same RIBONUCLEASE III, Dicer, that produces small interfering RNAs (RNA, SMALL INTERFERING). They become part of the RNA-INDUCED SILENCING COMPLEX and repress the translation (TRANSLATION, GENETIC) of target RNA by binding to homologous 3'UTR region as an imperfect match. The small temporal RNAs (stRNAs), let-7 and lin-4, from C. elegans, are the first 2 miRNAs discovered, and are from a class of miRNAs involved in developmental timing.Primed In Situ Labeling: A technique that labels specific sequences in whole chromosomes by in situ DNA chain elongation or PCR (polymerase chain reaction).Ribonuclease III: An endoribonuclease that is specific for double-stranded RNA. It plays a role in POST-TRANSCRIPTIONAL RNA PROCESSING of pre-RIBOSOMAL RNA and a variety of other RNA structures that contain double-stranded regions.Molecular Farming: The large scale production of pharmaceutically important and commercially valuable RECOMBINANT PROTEINS.Vaccines, Edible: Vaccines or candidate vaccines derived from edible plants. Transgenic plants (PLANTS, TRANSGENIC) are used as recombinant protein production systems and the edible plant tissue functions as an oral vaccine.Malaysia: A parliamentary democracy with a constitutional monarch in southeast Asia, consisting of 11 states (West Malaysia) on the Malay Peninsula and two states (East Malaysia) on the island of BORNEO. It is also called the Federation of Malaysia. Its capital is Kuala Lumpur. Before 1963 it was the Union of Malaya. It reorganized in 1948 as the Federation of Malaya, becoming independent from British Malaya in 1957 and becoming Malaysia in 1963 as a federation of Malaya, Sabah, Sarawak, and Singapore (which seceded in 1965). The form Malay- probably derives from the Tamil malay, mountain, with reference to its geography. (From Webster's New Geographical Dictionary, 1988, p715 & Room, Brewer's Dictionary of Names, 1992, p329)Coleus: A plant genus of the family Lamiaceae. The species of Coleus should be distinguished from PLECTRANTHUS BARBATUS - which is also known as Coleus forskohlii.NebraskaFellowships and Scholarships: Stipends or grants-in-aid granted by foundations or institutions to individuals for study.Biotechnology: Body of knowledge related to the use of organisms, cells or cell-derived constituents for the purpose of developing products which are technically, scientifically and clinically useful. Alteration of biologic function at the molecular level (i.e., GENETIC ENGINEERING) is a central focus; laboratory methods used include TRANSFECTION and CLONING technologies, sequence and structure analysis algorithms, computer databases, and gene and protein structure function analysis and prediction.Tetrahymena: A genus of ciliate protozoa commonly used in genetic, cytological, and other research.Tetrahymena pyriformis: A species of ciliate protozoa used extensively in genetic research.Tetrahymena thermophila: A species of ciliate protozoa used in genetic and cytological research.Electroporation: A technique in which electric pulses of intensity in kilovolts per centimeter and of microsecond-to-millisecond duration cause a temporary loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA.Transformation, Genetic: Change brought about to an organisms genetic composition by unidirectional transfer (TRANSFECTION; TRANSDUCTION, GENETIC; CONJUGATION, GENETIC, etc.) and incorporation of foreign DNA into prokaryotic or eukaryotic cells by recombination of part or all of that DNA into the cell's genome.Saccharomyces cerevisiae: A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.

Activation of systemic acquired silencing by localised introduction of DNA. (1/261)

BACKGROUND: In plants, post-transcriptional gene silencing results in RNA degradation after transcription. Among tobacco transformants carrying a nitrate reductase (Nia) construct under the control of the cauliflower mosaic virus 35S promoter (35S-Nia2), one class of transformants spontaneously triggers Nia post-transcriptional gene silencing (class II) whereas another class does not (class I). Non-silenced plants of both classes become silenced when grafted onto silenced stocks, indicating the existence of a systemic silencing signal. Graft-transmitted silencing is maintained in class II but not in class I plants when removed from silenced stocks, indicating similar requirements for spontaneous triggering and maintenance. RESULTS: Introduction of 35S-Nia2 DNA by the gene transfer method called biolistics led to localised acquired silencing (LAS) in bombarded leaves of wild-type, class I and class II plants, and to systemic acquired silencing (SAS) in class II plants. SAS occurred even if the targeted leaf was removed 2 days after bombardment, indicating that the systemic signal is produced, transmitted and amplified rapidly. SAS was activated by sense, antisense and promoterless Nia2 DNA constructs, indicating that transcription is not required although it does stimulate SAS. CONCLUSIONS: SAS was activated by biolistic introduction of promoterless constructs, indicating that the DNA itself is a potent activator of post-transcriptional gene silencing. The systemic silencing signal invaded the whole plant by cell-to-cell and long-distance propagation, and reamplification of the signal.  (+info)

Selective activation of heterologously expressed G protein-gated K+ channels by M2 muscarinic receptors in rat sympathetic neurones. (2/261)

1. G protein-regulated inward rectifier K+ (GIRK) channels were over-expressed in dissociated rat superior cervical sympathetic (SCG) neurones by co-transfecting green fluorescent protein (GFP)-, GIRK1- and GIRK2-expressing plasmids using the biolistic technique. Membrane currents were subsequently recorded with whole-cell patch electrodes. 2. Co-transfected cells had larger Ba2+-sensitive inwardly rectifying currents and 13 mV more negative resting potentials (in 3 mM [K+]o) than non-transfected cells, or cells transfected with GIRK1 or GIRK2 alone. 3. Carbachol (CCh, 1-30 microM) increased the inwardly rectifying current in 70 % of GIRK1+ GIRK2-transfected cells by 261 +/- 53 % (n = 6, CCh 30 microM) at -120 mV, but had no effect in non-transfected cells or in cells transfected with GIRK1 or GIRK2 alone. Pertussis toxin prevented the effect of carbachol but had no effect on basal currents. 4. The effect of CCh was antagonized by 6 nM tripitramine but not by 100 nM pirenzepine, consistent with activation of endogenous M2 muscarinic acetylcholine receptors. 5. In contrast, inhibition of the voltage-activated Ca2+ current by CCh was antagonized by 100 nM pirenzepine but not by 6 nM tripitramine, indicating that it was mediated by M4 muscarinic acetylcholine receptors. 6. We conclude that endogenous M2 and M4 muscarinic receptors selectively couple to GIRK currents and Ca2+ currents respectively, with negligible cross-talk.  (+info)

The DNA binding site of the Dof protein NtBBF1 is essential for tissue-specific and auxin-regulated expression of the rolB oncogene in plants. (3/261)

The Dof proteins are a large family of plant transcription factors that share a single highly conserved zinc finger. The tobacco Dof protein NtBBF1 was identified by its ability to bind to regulatory domain B in the promoter of the rolB oncogene. In this study, we show that the ACT T TA target sequence of NtBBF1 in domain B is necessary for tissue-specific expression of rolB. beta-Glucuronidase (GUS) activity of tobacco plants containing a rolB promoter-GUS fusion with a mutated NtBBF1 target sequence within domain B is almost completely suppressed in apical meristems and is severely abated in the vascular system. The ACT T TA motif is shown here also to be one of the cis-regulatory elements involved in auxin induction of rolB. The pattern of NtBBF1 expression in plants is remarkably similar to that of rolB, except in mesophyll cells of mature leaves, in which only NtBBF1 expression could be detected. Ectopic expression of rolB in mesophyll cells was achieved by particle gun delivery if the NtBBF1 binding sequence was intact. These data provide evidence that in the plant, a Dof protein DNA binding sequence acts as a transcriptional regulatory motif, and they point to NtBBF1 as the protein involved in mediating tissue-specific and auxin-inducible expression of rolB.  (+info)

Identification of cis-acting RNA leader elements required for chloroplast psbD gene expression in Chlamydomonas. (4/261)

The psbD mRNA of Chlamydomonas reinhardtii is one of the most abundant chloroplast transcripts and encodes the photosystem II reaction center polypeptide D2. This RNA exists in two forms with 5' untranslated regions of 74 and 47 nucleotides. The shorter form, which is associated with polysomes, is likely to result from processing of the larger RNA. Using site-directed mutagenesis and biolistic transformation, we have identified two major RNA stability determinants within the first 12 nucleotides at the 5' end and near position -30 relative to the AUG initiation codon of psbD. Insertion of a polyguanosine tract at position -60 did not appreciably interfere with translation of psbD mRNA. The same poly(G) insertion in the nac2-26 mutant, which is known to be deficient in psbD mRNA accumulation, stabilized the psbD RNA. However, the shorter psbD RNA did not accumulate, and the other psbD RNAs were not translated. Two other elements were found to affect translation but not RNA stability. The first comprises a highly U-rich sequence (positions -20 to -15), and the second, called PRB1 (positions -14 to -11), is complementary to the 3' end of the 16S rRNA. Changing the PRB1 sequence from GGAG to AAAG had no detectable effect on psbD mRNA translation. However, changing this sequence to CCUC led to a fourfold diminished rate of D2 synthesis and accumulation. When the psbD initiation codon was changed to AUA or AUU, D2 synthesis was no longer detected, and psbD RNA accumulated to wild-type levels. The singular organization of the psbD 5' untranslated region could play an important role in the control of initiation of psbD mRNA translation.  (+info)

Transient expression of DNA and RNA in parasitic helminths by using particle bombardment. (5/261)

Parasitic helminths (worms belonging to several metazoan phyla) cause considerable morbidity and mortality in humans. They are an important veterinary problem, and they result in significant economic losses in animal grazing and agriculture. Experimental studies on parasitic helminths have been limited by a lack of parasite cell lines and methods for molecular genetic analyses. We evaluated particle bombardment (biolistics) as a strategy to introduce and express nucleic acids in these multicellular parasites. By using embryos of the parasitic nematode Ascaris as a model, we developed methods to introduce and express both DNA and RNA during several stages of Ascaris embryogenesis. Biolistic transfection will facilitate experimental strategies in Ascaris embryos complementing other biochemical tools available (e.g., in vitro whole-cell embryo extracts for transcription, RNA processing, and translation). Transfection experiments with adult schistosomes further suggest that the biolistic strategy should be applicable to a variety of other parasitic helminths. The development of these methods provides molecular genetic tools to study gene expression and the biology of a variety of types and developmental stages of important helminth parasites.  (+info)

Co-delivery of T helper 1-biasing cytokine genes enhances the efficacy of gene gun immunization of mice: studies with the model tumor antigen beta-galactosidase and the BALB/c Meth A p53 tumor-specific antigen. (6/261)

DNA-based immunization is currently being investigated as a new method for the induction of cellular and humoral immunity directed against viral disease and cancer. In the present study we characterized and compared the immune responses induced in mice following particle-bombardment of the skin ('gene gun' immunization) with those elicited by intracutaneous injection of a recombinant adenoviral vector. Using the well characterized beta-galactosidase (beta gal) model Ag system we find that both in vivo gene transfer systems elicit potent and long-lasting anti-beta gal-specific CD8+ and CD4+ T cell responses. However, gene gun immunization predominantly promotes the production of anti-beta gal antibodies of the gamma 1 isotype, indicative of a Th2-biased immune response, while intradermal injection of recombinant adenovirus primarily leads to the production of anti-beta gal gamma 2a antibodies, indicative of a Th1-biased immune response. Since viral infections are generally associated with the production of large amounts of IFN-alpha and IL-12, we investigated whether administration of expression plasmids encoding these Th1-associated cytokines along with antigen-encoding cDNA can influence the nature of the immune response resulting from gene gun immunization. We observed that co-delivery of IFN-alpha or IL-12 resulted in increased production of anti-beta gal gamma 2a antibodies. This suggests a shift towards a Th1 phenotype of the resulting immune response, thus mimicking a viral infection. Importantly, gene gun immunization of mice with a naturally occurring tumor antigen, the tumor-specific p53 mutant antigen expressed by the chemically induced BALB/c Meth A sarcoma, required co-delivery of IL-12 for the induction of effective antitumor immunity. These results have important implications for the design of clinically relevant gene gun immunization strategies for tumor immunotherapy.  (+info)

Immune responses induced by gene gun or intramuscular injection of DNA vaccines that express immunogenic regions of the serine repeat antigen from Plasmodium falciparum. (7/261)

The liver- and blood-stage-expressed serine repeat antigen (SERA) of Plasmodium falciparum is a candidate protein for a human malaria vaccine. We compared the immune responses induced in mice immunized with SERA-expressing plasmid DNA vaccines delivered by intramuscular (i.m.) injection or delivered intradermally by Gene Gun immunization. Mice were immunized with a pcdna3 plasmid encoding the entire 47-kDa domain of SERA (amino acids 17 to 382) or the N-terminal domain (amino acids 17 to 110) of SERA. Minimal antibody responses were detected following DNA vaccination with the N-terminal domain of SERA, suggesting that the N-terminal domain alone is not highly immunogenic by this route of vaccine delivery. Immunization of mice by Gene Gun delivery of the 47-kDa domain of SERA elicited a significantly higher serum antibody titer to the antigen than immunization of mice by i.m. injection with the same plasmid did. The predominant isotype subclass of the antibodies elicited to the SERA protein following i.m. and Gene Gun immunizations with SERA plasmid DNA was immunoglobulin G1. Coimmunization of mice with SERA plasmid DNA and a plasmid expressing the hepatitis B surface antigen (pCMV-s) by the i.m. route resulted in higher anti-SERA titers than those generated in mice immunized with the SERA DNA plasmid alone. Vaccination with DNA may provide a viable alternative or may be used in conjunction with protein-based subunit vaccines to maximize the efficacy of a human malaria vaccine that includes immunogenic regions of the SERA protein.  (+info)

Effective DNA vaccination against listeriosis by prime/boost inoculation with the gene gun. (8/261)

Protective immunity against Listeria monocytogenes strongly depends on CD8+ T lymphocytes, and both IFN-gamma secretion and target cell killing are considered relevant to protection. We analyzed whether we could induce a protective type 1 immune response by DNA vaccination with the gene gun using plasmids encoding for two immunodominant listerial Ags, listeriolysin and p60. To induce a Th1 response, we 1) coprecipitated a plasmid encoding for GM-CSF, 2) employed a prime/boost vaccination schedule with a 45-day interval, and 3) coinjected oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs. DNA immunization of BALB/c mice with plasmids encoding for listeriolysin (pChly) and p60 (pCiap) efficiently induced MHC class I-restricted, Ag-specific CD8+ T cells that produced IFN-gamma. Coinjection of CpG-ODN significantly increased the frequency of specific IFN-gamma-secreting T cells. Although pChly induced specific CD8+ T cells expressing CTL activity, it failed to stimulate CD4+ T cells. Only pCiap induced significant CD4+ T cell and humoral responses, which were predominantly of Th2 type. Vaccination with either plasmid induced protective immunity against listerial challenge, and coinjection of CpG ODN improved vaccine efficacy in some situations. This study demonstrates the feasibility of gene gun administration of plasmid DNA for inducing immunity against an intracellular pathogen for which protection primarily depends on type 1 CD8+ T cells.  (+info)

*Agricultural biotechnology

New genetic information can be inserted with biolistics. An example of transgenics is the rainbow papaya, which is modified ...

*Gene gun

Biolistics introduces DNA randomly into the target cells. Thus the DNA may be transformed into whatever genomes are present in ... Biolistics has proven to be a versatile method of genetic modification and it is generally preferred to engineer transformation ... Notably, Bt maize is a product of biolistics. Plastid transformation has also seen great success with particle bombardment when ... John O'Brien presents...Gene Gun Barrels for more information about biolistics. ...

*Genetic engineering techniques

Due to the associated cell and DNA damage, the transformation efficiency of biolistics and electroporation is lower than with ... In plants the DNA is generally inserted using Agrobacterium-mediated recombination or biolistics. In Agrobacterium-mediated ... This can be accomplished with microinjection, viral vectors, Agrobacterium-mediated recombination, biolistics or using the ...

*Plant genetics

The "Gene Gun" method is also referred to as "biolistics" (ballistics using biological components). This technique is used for ...

*Genetically modified tomato

Genetic material can also be inserted into a tomato cell's chloroplast and chromoplast plastomes using biolistics. Tomatoes ...

*Triticale

... has, until recently, only been transformed via biolistics, with a 3.3% success rate (Zimny et al. 1995). Little has ... Two methods are commonly employed: infectious bacterial-mediated (usually Agrobacterium) and biolistics, with the latter being ...

*Transformation (genetics)

... or biolistics. Particles of gold or tungsten are coated with DNA and then shot into young plant cells or plant embryos. Some ...

*Transgene

To combat this, scientists used biolistics to insert the daffodil phytoene synthase gene into Asia indigenous rice cultivars. ... biolistics which is the procedure of shooting DNA bullets into cells, and also delivering DNA into the egg that has just been ...

*Genetically modified crops

Gene guns (also known as biolistics) "shoot" (direct high energy particles or radiations against) target genes into plant cells ...

*Genetic engineering

Other methods include biolistics, where particles of gold or tungsten are coated with DNA and then shot into young plant cells ... Due to the damage caused to the cells and DNA the transformation efficiency of biolistics and electroporation is lower than ...
... ,The Helios gene gun low-pressure system, 100-120 V, is used for biolistic particle delivery of biomaterials into cells. The system includes the Helios gene gun, helium hose assembly, helium regulator, tubing prep station, syringe kit, tubing cutter, Helios gene gun optimization kit, low-pressure re,biological,biology supply,biology supplies,biology product
Retinas were isolated from Brown Norwegian male rats 9-12 weeks of age and cultured on 0.4 µm Millicell culture plate inserts in Neurobasal-A medium at 37ºC in 5% CO2. A Helios gene gun system was used for particle-mediated transfer of a YFP-expressing plasmid to RGCs. Retinal explants were treated with a broad-spectrum, irreversible caspase inhibitor (Boc-D-FMK) to prevent apoptosis. The morphology of RGCs was observed by confocal microscopy. ...
DNA can be stably targeted and integrated in the somatic or germline genomes of Tetrahymena using homologous recombination. The DNA cassettes described below enable gene knockouts, knockins, and mutations.. Transformation of DNA into Tetrahymena is performed using microinjection, electroporation, or biolistics. Transformations require little DNA (microgram quantities) and positive transformants are acquired in ,1 week. Transformation of DNA into the MAC or the MIC depends on the method used; in microinjections, needles can specifically deliver DNA to the MAC, whereas electroporation and biolistics rely on the precise timing of the transformation during the cell cycle or during conjugation. Recovery of MIC transformants requires mating, while MAC transformants can be selected directly. Transformations using biolistics are simple and efficient, making it the method of choice. This is especially important for low-efficiency transformations that target MIC DNA integration (Cassidy-Hanley et al. ...
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4Embrapa Suínos e Aves, Concórdia, SC, Brasil. ABSTRACT Fertilized chicken eggs were bombarded with a biolistic device. Transient expression of the lacZ gene under the control of a human cytomegalovirus (CMV) promoter was assessed after in situ gene transfer using this approach. The influence of different pressures, vacuum levels and particles was tested. Survival rate improved as particle velocity decreased, but resulted in lower levels of expression. The best survival and expression were obtained with gold particles, a helium gas pressure of 600 psi and a vacuum of 600 mmHg. Under these conditions, all bombarded embryos showed b-galactosidase activity, indicating that this was an effective method for transformation of chicken embryos. INTRODUCTION DNA transfer methods have led to significant advances in exogenous gene expression. Transgenic animals have been used as models for studying gene functions and for producing very marketable proteins. Microinjection has been the most studied form of ...
A gene gun or a biolistic particle delivery system, originally designed for plant transformation, is a device for injecting cells with genetic information. The payload is an elemental particle of a heavy metal coated with plasmid DNA. This technique is often simply referred to as bioballistics or biolistics ...
Endpoint immunoglobulin G (IgG) titers and cytotoxic T-lymphocyte (CTL) activities were identical between mice immunized via the intramuscular and epidermal (gene gun) routes with 100 and 1 micrograms, respectively, of an influenza virus nucleoprotein (NP) expression vector. However, examination of the relative levels of two IgG subclasses demonstrated that muscle inoculation resulted in predominantly IgG2a responses, whereas gene gun immunization yielded a preponderance of IgG1 antibodies. Inasmuch as these data suggested that muscle inoculation and gene gun delivery elicited Th1-like and Th2-like responses, respectively, gamma interferon release profiles from antigen-stimulated splenocytes were remarkably similar between these groups. Interleukin-4 (IL-4) production assays, on the other hand, revealed qualitative differences that could be correlated with the divergent IgG subclass data. Waning gamma interferon production in gene gun-immunized animals was countered by a marked increase in IL-4 ...
...Peter Jackstadt 1 Horst Zahner 2 and Gerd Hobom ... Introduction The Biolistic DNA transf...Here we report on the potential to use this convenient method for tran...,Transformation,of,Nematodes,With,the,Helios,Gene,Gun,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology
Transgenic trees currently are being produced by Agrobacterium-mediated transformation and biolistics. Since trees are particularly suited for long-term evaluations of the impact of the technology, Prunus subhirtella autumnorosa (PAR) was chosen as model fruit tree species and transformed with a reporter gene (uidA) under the control of the 35S promoter. Using Southern and GUS fluorometric techniques, we compared transgene copy numbers and observed stability of transgene expression levels in 34 different transgenic plants, grown under in vitro, greenhouse and screenhouse conditions, over a period of 9 years. An influence of grafting on gene expression was not observed. No silenced transgenic plant was detected. Overall, these results suggest that transgene expression in perennial species, such as fruit trees, remains stable in time and space, over extended periods and in different organs, confirming the value of PAR as model species to study season-dependent regulation in mature stonefruit ...
Cellular introduction of PEBBLEs (photonic explorers for bioanalysis with biologically localized embedding) has been investigated by a wide variety of methods in a range of cell types. These methods include surface functionalization with CPPs (cell-penetrating peptides), pinocytosis, commercial lipid transfection agents, cytochalasin D, picoinjection, and Gene gun bombardment. This paper will overview several of the most popular methods used for the introduction of PEBBLE nanosensors to the cellular environment and discuss the efficacy of the techniques.. ...
Intravenously injected nanopharmaceuticals induce adverse cardiopulmonary reactions in sensitive human subjects and these reactions are reproducible in pigs. The underlying mechanisms are poorly understood, but a role for both the complement system and reactive macrophages has been implicated. Here we show the dominance and importance of early pulmonary intravascular macrophage clearance kinetics in adverse particle-mediated cardiopulmonary distress in pigs and irrespective of complement activation. Delaying particle recognition by macrophages within the first few minutes of injection overcome adverse reactions in pigs. This was achieved by two independent approaches: (i) changing particle geometry from a spherical shape (which trigger cardiopulmonary distress) to either rod- or disk-shape morphology and (ii) by physically adhering spheres to the surface of erythrocytes. These approaches bypasses particle surface engineering approaches to prevent robust macrophage recognition as well as the use ...
We tested this hypothesis through HvWRKY2 overexpression experiments during compatible interactions. Biolistic delivery of a HvWRKY2 construct, driven by the strong ubiquitin promoter, into single leaf epidermal cells resulted in supersusceptibility in different genetic backgrounds harboring MLA1-HA, MLA6-HA, or wild-type MLA10 (Fig. 3B; similar results were obtained with HvWRKY1). Overexpression of SUSIBA2, a barley WRKY TF functioning in sugar signaling (15), did not alter the B. graminis infection type (Fig. 3B), indicating that sequence motifs other than the shared WRKY DNA binding domain (16) contribute to the HvWRKY1/2-dependent supersusceptible phenotype. The contrasting infection phenotypes observed upon overexpression or gene silencing of HvWRKY1/2 are consistent with their presumed roles as repressors of basal defense. HvWRKY1 and HvWRKY2 expression was strongly (≥20 fold), rapidly (within 3 hours), and transiently activated upon B. graminis challenge in both compatible and ...
Modern transformation and genome editing techniques have shown great success across a broad variety of organisms. However, no study of successfully applied genome editing has been reported in a dinoflagellate despite the first genetic transformation of Symbiodinium being published about 20 years ago. Using an array of different available transformation techniques, we attempted to transform Symbiodinium microadriaticum (CCMP2467), a dinoflagellate symbiont of reef-building corals, in order to perform CRISPR-Ca9 mediated genome editing. Plasmid vectors containing the chloramphenicol resistance gene under the control of the CaMV p35S promoter as well as several putative endogenous promoters were used to test a variety of transformation techniques including biolistics, electroporation, silica whiskers and glass bead agitation. We report that we have been unable to confer chloramphenicol resistance to our specific Symbiodinium strain. These results are intended to provide other researchers with an ...
The presence of foreign genes that were transferred into Citrus nucellar cells by the particle bombardment system has been expressed with the GUS assay. Tungsten microcarriers coated with plasmid DNA containing the gus A gene (pBI221.2) were accelerated at high velocity using a biolistic device into lemon (Citrus limon var. Kütdiken) target cells. Histochemical examination of the bombarded nucellar cells revealed that only several cell aggregates had cells expressing the gus A gene. Control cell aggregates that were not bombarded did not exhibit any cells expressing the gus A gene.. ...
The spleens of intramuscularly or gene gun immunized mice enlarged notably compared with those of the unimmunized controls. In particular, the spleen weight of the gene gun injected mice almost doubled that of the unimmunized controls (data not shown). FACS analysis of spleen cells harvested 2 weeks after the final immunization with AMA-1 or AMA-1 plus IL-12 DNA vaccines showed notable differences in the proportions of CD4+ and CD8+ T-lymphocytes between the intramuscular injection versus the gene gun (epidermis) injection (Fig. 2). In mice immunized intramuscularly (n=5 for each group) with AMA-1 alone or AMA-1 plus IL-12 DNA vaccines, the proportions of T-cell subsets (17.9±0.47% or 15.5±0.79% for CD8+ T-cells and 27.6±0.84% or 27.7±0.72% for CD4+ T-cells) did not show any significant changes (P,0.05), compared with control mice immunized only with UB vector (16.2±0.78% for CD8+ T-cells and 26.5±0.35% for CD4+ T-cells) (Fig. 2). By contrast, in mice injected with a gene gun (n=3 for each ...
Agrobacterium-mediated transformation and direct gene transfer using the gene gun (microparticle -bombardment) are the two most widely used methods for plant genetic modification. The Agrobacterium method has been successfully practiced in dicots for
Dendritic cells, the security cameras of the immune system, derive their name from their finger-like projections. They continually capture external proteins, digest the proteins into fragments, and display those fragments on their surfaces. T cells, the police who watch the cameras, have the ability to examine the fragments on the dendritic cells surfaces and sound the alarm to the rest of the immune system if they determine that those fragments are dangerous. Although other kinds of cells also have the ability to present fragments of foreign proteins to the immune system, dendritic cells are the most proficient, and immunologists call them "professional" antigen-presenting cells ...
Cell culture, transfection, immunocytochemistry, and Western blot. Calcium phosphate transfections, immunocytochemistry, and culture of dissociated hippocampal CA3/CA1 pyramidal neurons were performed as described previously (Wu et al., 2001). Enhanced green fluorescent protein (EGFP) was cotransfected with hemagglutinin (HA)- or myc-tagged constructs to visualize the detailed cell morphology. Hippocampal slice culture was done from 7-8 d postnatal rats and transfected biolistically at 2 d in vitro (DIV) as described previously (Lo et al., 1994). A modified procedure was used for Western blot experiments using cells growing on 12 mm coverslips. In brief, coverslips were placed in 1.5 ml microcentrifuge tubes containing 40 μl of 9 m urea and were crushed, and, after vigorous vortexing, samples were centrifuged (18,000 × g for 2 min). The protein concentration was determined by BCA assay (Pierce, Rockford, IL), and volumes were adjusted with 9 m urea so that all samples were 0.3-1.0 μg/μl. ...
In a new study, Debra Hansen, a research professor at Arizona State Universitys Biodesign Institute, explores an innovative means of investigating membrane proteins produced by a pair of highly pathogenic organisms. The research team showed that DNA-based genetic immunization, using a device known as a gene gun, could successfully express membrane proteins in mice and induce the animals to produce a range of critical antibodies to bacterial and viral targets.
The first time I read the term biolistic, I thought it was a misprint. Then I found out it was a chimera-like word, with a "biological" head and a "ballistic" body. The term describes a method for introducing DNA into cells by literally shooting them with microscopic bullets, which have been previously coated with the desired DNA. Although the technique is mainly used for the genetic engineering of plants, sometimes it is also employed for animal cells and tissues, fungi or bacteria. I have never used a gene gun, not even been close to one, so I greatly enjoyed the following video from JoVE (Journal of Visualized Experiments, a video journal for biological research, see note* ...
The gene gun worked with a broader range of plant crops than agrobacterium in the early days and so it was a more random method, it was also more broadly useful. Today, agrobacterium has been adapted to plants that previously it was thought not to work on, and I think in the long run most engineering will go back to the agrobacterium system ...
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It is usually better to be a contrarian, doing something that makes you stand out from the crowd. A gun promotion fits this criteria, giving the marketer significant publicity for free. In addition, the popularity of guns has been downplayed by the Establishment media. This gives those who go against the narrative that "guns are bad" an advantage, because they appeal to the large majority of Americans who believe, contrary to the narrative, that guns are valuable, and thus, good ...
Spaceship guns. Guns can only exist in Life-like rules that include the B2 or B3 rule. To the best of my knowledge, this mini-collection includes all of the rules with B3 for which a spaceship gun is known. (Im not attempting to collect every gun known; Im just trying to include at least one gun for every rule in which a gun is known.) In some rules with B2, guns with a period as low as 4 can exist. Periods that low can be searched for directly using oscillator search programs, making them potentially much easier to find. However, the low period combined with the explosiveness of B2 rules makes it difficult build anything interesting out of these guns. This collection does not necessarily contain a gun for every B2 rule in which a gun is known. [Download all the guns in a single ZIP file: alienguns.zip] B3/S23 - (Life) p44 MWSS (c/2 orthogonal ...
In trying to bridge the civic divide over guns, a productive approach to conversation is to focus not on who is right, but on where people are coming from. Questions developed at a recent public forum on guns, hosted by The Monitor, can be tools for less polarized dialogue on guns.
Just because you mention "beaten to death", doesnt necessarily mean it refers to the gun, the knife, or the improvised weapon. Poor syntax. And no, i - #113026427 added by comradewinter at Guns Arent The Problem
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While little is officially known about Calamity Jane, one thing is for sure: her nickname certainly seems to fit what we do know about her.
DNA vaccines expressing plasma membrane and secreted forms of the influenza and measles virus hemagglutinins (HAs) have been used to evaluate the effect of secretion on DNA-raised antibody responses. At low doses of DNA, the plasma membrane form of the influenza virus HA raised higher titers of antibody than the secreted form. The isotype of the DNA-raised antibodies depended on both the method of DNA delivery and the form of the expressed antigen. Following intramuscular injections, DNAs expressing membrane bound forms of the influenza and measles HAs raised predominantly IgG2a. By contrast, DNAs expressing the secreted from of the two HAs as well as another secreted protein, human growth hormone, raised predominantly IgG1. Gene gun delivery resulted in predominantly IgG1 antibody responses for both secreted and membrane bound forms of the hemagglutinins. The raising of predominantly IgG1 by i.m. delivery of the secreted form of the influenza hemagglutinin was IL-4 dependent suggesting that a T-helper
Principal Investigator:UMEHARA Takashi, Project Period (FY):1999 - 2000, Research Category:Grant-in-Aid for Scientific Research (C), Section:一般, Research Field:Orthopaedic surgery
Slices and electrophysiology. All experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH publication no. 86-23; revised 1987). Cultured slices were prepared from the primary visual cortex of postnatal day 0 (P0)-P1 C57 mice as described (Dunaevsky et al., 1999), incubated for 2 weeks, and biolistically transfected with cytomegalovirus-EGFP vector (Clontech, Palo Alto, CA). After 2-4 d, slices were imaged in artificial CSF (ACSF) that contained (in mm): 126 NaCl, 3 KCl, 2 CaCl2, 2 MgSO4, 1.1 NaH3PO4, 26 NaHCO3, and 10 dextrose, bubbled with 95% O2 and 5% CO2. Acute slices were made from primary visual cortex from P12 to P16 mice as described (Peterlin et al., 2000). Layer 2/3 pyramidal neurons in V1 were selected under differential interference contrast, and whole-cell recordings were made with an Axoclamp 2B (Axon Instruments, Foster City, CA) amplifier. Borosilicate pipettes with an outer diameter of 1.5 mm and an ...
... : Particle bombardment of coat protein into epidermal cells. Coat protein was cloned in fusion to green fluorescent protein into pRT101- transient expression plasmid under control of CaMV 35S promoter and introduced into epidermal cells by biolistic means (A). Coat protein was extracted from bombarded leaves and detected using TMV-specific antiserum (B). M. Prestained Protein Marker, Broad Range (New England BioLabs, UK), 1. wild-type tobacco, 2. CP-GFP from bombarded leaves ...
Immature scutella of barley were transformed with cDNA coding for a 13-li-poxygenase of barley (LOX-100) via particle bombardment. Regenerated plants were tested by PAT-assay, Western-analysis and PCR-screening. Immunocytochemical assay of T0 plants showed expression of the LOX cDNA both in the chloroplasts and in the cytosol, depending on the presence of the chloroplast signal peptide sequences in the cDNA. A few transgenic plants containing higher amounts of LOX-derived products have been found. These are the candidates for further analysis concerning pathogen resistance ...
Immature scutella of barley were transformed with cDNA coding for a 13-li-poxygenase of barley (LOX-100) via particle bombardment. Regenerated plants were tested by PAT-assay, Western-analysis and PCR-screening. Immunocytochemical assay of T0 plants showed expression of the LOX cDNA both in the chloroplasts and in the cytosol, depending on the presence of the chloroplast signal peptide sequences in the cDNA. A few transgenic plants containing higher amounts of LOX-derived products have been found. These are the candidates for further analysis concerning pathogen resistance ...
There is no reason why you cant attach the peptide to a protein carrier such as KLH or ovalbumin to increase the immunogenicity. Yes, you will get antibodies to the protein as well, but you need to make sure your screening assays are specific for the peptide, you should be okay. Another suggestion, try hamsters instead of rats. DNA is also an option, although the gene gun approach seems to be more successful than im injections of DNA. -ned Daniele Focosi ,mi at interhealth.info, wrote in message news:e794ccc8.0209111128.5ddb060b at posting.google.com... , I would like to suggest you to prove DNA immunization for better , results. Anyway please note Im just a PhD student and I have not real , experience with such a technique apart from literature results ! , Please let me know. , Best regards. , , Daniele , , news.dk.uu.net ,myd at nordicbioscience.com, wrote in message news:,3d78af5a$0$10685$4d4eb98e at news.dk.uu.net,... , , Dear all, , , , , I would like to know if it is possible to ...
PLEASE READ,. I bought this gun out of impulse. I was looking for a Kel-Tec P3AT. Recently moved to Indiana where buying a gun is so much easier than in Massachusetts. After searching for the Kel-Tec I found that it was not within my very small budget and without reading reviews I bought the Taurus PT 738 TCP .380 acp. After my purchase (which I never recommend. Always do your research first) I decided to look up reviews. To my luck (Im never this lucky) I read some excellent reviews. I was excited for this new purchase and then….. I came across some not so good ones. In fact, some people even trashed the gun. Some quick background. I have never owned a gun. Prior to holding my new gun I probably held a gun twice in my life and never had fired one. When I got my Taurus it was the first time I had fired it. In comparison to shooting and the ergonomics to other manufacturers I cannot say. What I can say is that I have already put nearly 250 rounds to this baby and boy has it lived up to all the ...
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A perforating gun gravitational orientation system includes a perforating gun and a swivel device connected to the perforating gun to permit rotation of the perforating gun within casing, and the perf
Texas Gun Talk is the most active and largest Texas gun forum on the net. Be sure to check out our gun classifieds, political, and hometown sections.. ...
Mrs. Lanza has a right to own a gun. However, that doesnt guarantee that shell be able to find any gun that her little heart desires available for purchasing in the U.S. nor does that allow her to purchase a weapon without jumping through hoops. I want the hoops to be many. Here are the new rules:. 1) There will be a gun permit required to purchase ANY gun in NYS - long gun, handgun, whatever now is defined as a GUN. Obviously, that does NOT include paintball equipment or airguns used by kids to hunt squirrels etc. To get a gun purchasing permit, you MUST be over 18 and submit to the background check and appearance before a judge thats currently required for a handgun. There will be a significant waiting period. The current fee will NOT be raised. 2)You have five years to obtain a gun permit for any existing gun that you own. In the meantime, the gun cannot be taken outside of your legal residence. Period. The State of New York WILL provide sufficient judges (perhaps administrative ones) so ...
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Law-abiding gun owners have reason to be very concerned by the recent actions of the Trudeau Liberals. While in government, Conservatives made a promise to scrap the wasteful and ineffective Long-Gun Registry – and we kept that promise. Now the Liberals are doing everything in their power to create a back-door registry that will allow […]
ဒါေပမဲ့ ေဒလီမွာ Toy gun ကေလးကစား ေသနတ္တလက္ ဝယ္လာတာကို ကိုဗလ (သူလဲ ႏိုင္ငံေရးမွာ လုပ္ေဖၚကိုင္ဖက္ဘဲ) ေရွ႕မွာ ေနာက္ျပန္လွည့္ျပီး၊ နံရံေပၚက ပိန္ေညာင္ကို ျပစ္ျပတာ ဒက္ဒိမွန္သြားေတာ့ ကိုဗလခင္ျမာ ဆရာ့ကို ဘရာဗို မလုပ္ဘဲေနလို႔မရ ျဖစ္သြားတယ္ ...
The Guns of Navarone is a novel by Alistair MacLean, published in 1957, and more famously a classic 1961 film adaptation written by Carl Foreman and …
Hello all, 1) chainsaw 2) pancor jackhammer 3) m16 with undercarriage grenade launcher (arnolds gun in Predator or something similar to that) 4) minig...
558855 KG-3 is ideal for use after a coat of the KG-1 and KG-2 (sold separately). Also works to clean and degrease parts by rapidly cutting through oils...
hey if any1 would like me to make them a signature or like an avator just let me no, my sig below i did, and i also did panther 88 so i can do something like that for u if ud like ...
I learned something today. Something really important. About how to stick to your guns, how use your gut. About what it really means to be a doctor.
Genetic vaccination using naked plasmid DNA is an immunization strategy both against infectious diseases and cancer. In order to improve the efficacy of DNA vaccines, particularly in large animals and humans, different strategies have been pursued. These vaccination strategies are based on different application routes, schedules, and coexpression of immunomodulatory molecules as adjuvants. Our mouse tumor model offers the possibility to investigate Her2/neu DNA vaccines in different settings, i.e., intramuscular or intradermal application with or without coexpression of adjuvants. Protection from tumor growth in tumor challenge experiments and both T cell and humoral immune responses against Her2/neu peptides are used as surrogate parameters for vaccine efficacy.. ...
As a cytokine with functions directed to inflammatory and immune cells as well as fibroblasts, interleukin-11 was selected to modulate invasive behaviour of fibroblasts. For transduction, a retroviral vector system, particle-mediated transfer (gene gun), and a lentiviral vector based on simian immunodeficiency virus were investigated. Cells (500,000) were injected directly into the knee joinst of mice with severe combined immunodeficiency (SCID). Mice were monitored for joint swelling, serological parameters and by radiological methods. In vitro, cells were coincubated with cartilage for 1 week. Finally, the histology of cartilage destruction and signs of apoptosis were studied. ...
This object is a sample of tobacco leaf cell colonies that have been successfully genetically transformed using the biolistic gene gun prototype produced by John Sanford, Ed Wolf, and Nelson Allen at Cornell University in Ithaca, New York. Biolistic gene guns are used to genetically transform plants by shooting microprojectiles (tiny bullets) covered in DNA into plant cells. The blue color of the cells is due to one of the genes that was inserted, the GUS reporter system. Cells in which the GUS gene was successfully taken in and utilized produced a blue chemical. The blue cells therefore represented the success of the genetic transformation via the biolistic process ...
Giza (الجيزة al-Gīza) is a Governorate to the west of the Egyptian capital Cairo - a city in its own right, but for a long time now absorbed as part of the heavily-populated and sprawling Cairo metropolis. Giza is best known as that part of Cairo closest to the world-famous Pyramids of Giza, situated high on the desert plateau immediately to the west of the urban district, itself located in the valley and centred around the Pyramids Road, linking central Cairo with the ancient wonders. One of the premier attractions of Egypt, if not the world, the Pyramids of Giza represent the archetypal pyramid structures of ancient Egyptian civilisation and - together with the Sphinx at the base of the Giza plateau - are the iconic image of Egypt.
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There is a broad range of methods that are employed in the process of producing GMOs. This often involves insertion of a gene of interest into living organisms depending on the species that you are working on. In plants mainly, two most common biotechnology-based techniques include; Agrobacterium-mediated transformation and bombardment of particles. According to the regulations given by FSANZ, it is a requirement that clear description of the method employed in genetically engineering plants is given.. Case study- Roundup Ready soy. This was produced using the particle bombardment method. This process of biotechnologically engineering soybeans involved; bombardment of the plant cells with microscopic particles of gold coated with DNA that contains the gene of interest. The gene of interest is the EPSPS gene that is derived from Agrobacterium. The aim of this is to introduce the novel gene of interest through the cell wall so that it integrates into the genetic material of the soy plant. The new ...
Filling the glue guns butane tank is easy. Just invert the gun, slip the butane canisters nozzle into the valve on the bottom of the gun and pump. (We were surprised that there was hardly any smell emitted from the gun or canister when fueling the gun.) What we werent sure about was when the guns tank was completely full, although the instructions said the indicator was when the gas "overflow[s] from the nozzle." (Not comforting, but whatever.) Instructions also said that we were to wait a few minutes for the gas to stabilize before attempting to light the gun, which we did. To ignite the gun, we pushed the fuel flow control (the wheel at the top of the grip) up and then turned it to the right until we heard the familiar hissing sound of gas. A quick pull of the ignition switch just under and back from the tip of the gun and the gun was on. Master claims that the gun will reach full heat at 3 to 4 minutes, but ours was ready in about 2 minutes.. For a cordless glue gun, the PortaPro has ...
The techniques of plant transformation are discussed. They are bacterial-mediated, electroporation transformation, biolistic (particle bombardment), and microbeam laser. By using different strategies, foreign gene(s) from various organisms can be inserted into plant genome and expressed. Bacteria-mediated transformation uses Agrobacterium as a vector to insert genes of interest. The other three directly deliver the plasmid into the plant. While Bacteria-mediated method is relatively simple, the other three can transform both monocot and dicot plant, and use various plant tissues as target cells. Public concerns about the impact that transgenic plant might have on the environment have partly slowed down the large-scale use of transgenic plants. These concerns are discussed. Some suggestions regarding the releasing of the transgenic plant under field conditions are also discussed.
Comprare Levitra in Svizzera è facile e puoi avere nel giro di pochi giorni i farmaci migliori per il trattamento della disfunzione erettile, repeatedly spiropent clenbuterol senza bisogno di recarti dal medico e senza presentarti in farmacia imbarazzato? Study Protocol for a Randomised Controlled Trial? (1990) costively go Science 247:1465-1468) or particle bombardment (see eg, Cheng, L! [5] A esso ha fatto seguito nel 2009 lEP The Fame Monster, avodart kaufen dal quale sono stati estratti i singoli Bad Romance, Telephone e Alejandro. Vi divennero venne provinciale decori mentre altre paludose venne assoluto. Pero yo aconsejo tomarse un relajante y ponerse una buena cantidad de crema anestésica, avodart buy ni te vas a enterar. Preventive measures for colonization of hospitalized patients with C! Supreme Court affirmed by an equally divided court a Court of Appeals ruling that it is constitutional to impose a different physical presence requirement for unmarried US? Changes in glucose ...
4844 Tie-2 stabilises pericyte/endothelial interactions during angiogenesis and is over expressed on tumor endothelium. A vaccine that targets endothelium over-expressing Tie-2 may result in vessel damage and stimulate an inflammatory cascade resulting in disease regression. To validate the use of Tie-2 as a target, human Tie-2 was used as a xenogenic vaccine in mice. Preliminary studies suggest that immunisation of balb/c mice by gene gun with DNA from the extracellular region of Tie-2 resulted in cessation of tumor growth. To determine if we could design a syngeneic vaccine that would work in both mice and humans a conserved region of Tie- 2 was cloned. HLA-A*0201 transgenic mice were immunised with this construct to determine if a repertoire of HLA-A*0201 T cells exist that recognise Tie-2. Within the Tie-2 region an HLA-A*0201 epitope was identified that is identical between mice and humans. Anchor modification of this epitope, increased binding to MHC. Immunisation of HLA-A*0201 mice with ...
In the micro view of evolutionary patterns, individuals may survive to produce gametes, some of which combine into zygotes that themselves develop into reproductively viable individuals which can disperse to new habitats. The machinery underlying this picture involves heritable characters that are transmitted from parents to offspring through DNA in chromosomes and organelles. This information is translated via RNA into proteins. Gametes can include novel DNA sequences formed by recombination of parental chromosomes, and/or by mutation. Microevolution can be thought of as small scale motion through an adaptive landscape, with a tendency for populations to get "trapped" at local fitness maxima.. Genetic engineering is a term encompassing many new technologies, such as plasmid or viral transformation of bacteria, gene insertion (gene guns, vectors etc.) into crop plants, site-directed mutagenesis, and gene therapy. Potential applications of these include improvements on input and output sides of ...
DNA immunization, also known as gene immunization, polynucleotide vaccine and DNA vaccine, is a newly established immunological theory and technique first discovered in the 1990s. Compared with current protein vaccine, DNA immunization is safer and more inexpensive. Besides, it induces a more effective immune response and is easy to be prepared. With the potential application in some specific field, DNA immunization is not only been widely used in anti-virus, bacteria, fungi, parasites as well as other anti-infection immunity, but also plays a n important role in tumor immunology and autoimmune disease. Like all other groups, Creative Biolabs follows up with the updated technique in DNA immunization, and made their own innovation by putting DNA immunization in the production of antibody. ...
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Yes, could take one apart and duplicate that, within the capabilities of whatever CNC machine youre using.. And… then youd have a duplicate of that ONE Python, which might have been fitted up just a little differently than your Python, or my Python, or that guy over there who owns a Python.. The thing about hand-fitted guns and gun parts is that it is very rare that two guns are exactly identical.. The first thing youll run into is that parts in the current market might not work in the CNC duplicated Python. The next thing you might find out is that perhaps the example you chose was slightly tighter or looser than you might have ideally engineered if you stood back and did it from the ground up, working out how to make the parts wear in evenly, etc.. Taking apart a gun with a looser fit and duplicating it is easier than taking apart a gun that is fit up very tightly and duplicating it. Look at some of the issues on some CNCed 1911s, where people think "Oh, well just tighten this up here, ...
The SNCF is a European leader in the production of passenger trains, high speed trains, metros and busses. During the yearly Finishing Road Show, the Graco truck made a stop at one of SNCFs facilities in Rouen. Here, Graco showed its latest product innovations such as the ProMix™ 2KS, our advanced proportioning system, the Pro Xp™ electrostatic gun range and the Mini-Merkur™ ES.. It was the Mini-Merkur ES that caught the attention of the SNCF as a possible solution to replace the cup guns used to paint small parts of the TGV. After the Finishing Road Show, a test with the Mini-Merkur ES in combination with G40™ Air-Assisted Airless guns was organized by the local Graco distributor, Clid Industrie, at the SNCF in Rouen. The test included product compatibility, spray quality and material consumption.. The results of the test showed that the Graco solution was more effective for the painters. The G40 gun offers more operator comfort and a higher productivity with Air-Assisted Airless ...
I was very pleased with this gun. I recieved it only yesterday so i can not say how long it lasts but i can say that the quality is very high. Echo1 only makes the best guns and i would qualify them as on of the best AEG manufactures out there. Now just because this is the polymer version its cheap plastic, no! I have a very hrd time telling the difference between the polymer and the metal on the gun. Even if the body is polymer all of the internals are metal. As for how it shoots, amazing. I was looking for about 400 fps when buying it so i got a tight bore barrel and a m120 spring (which i cant instal until the warrenty runs out) but i dont even need them. When i bought this gun inwas looking for a cqc gun with a high fps and lots o metal. I found hat when i bought the xcr-c. Another advantage is airsoft GIs point system. I was able to get a gun bag with this gun for free. Ive heard that the mags get wonly on this gun. I have not experienced that yet but am watching closley. If your mags ...
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Now that Borderlands 3 is out in the wild, we've been able to take a hard look at the game's new gun generation systems and analyze our results. Obviously, Borderlands 3 is built on the foundation of randomly generated weapons, so we can't just list the best ones we found and hope you find them too. To account for randomness, we've listed our best traits to look for in guns and highlighted some common stuff to look out for.
The feud between Machine Gun Kelly and Eminem is still going strong. And Machine Gun Kelly isnt backing down anytime soon, especially not after being the subject of Eminems latest diss track
STOCKTON - Three people were arrested after a routine traffic stop led authorities to a large amount of drugs, money and guns, police said.
If you structure a class correctly, you dont have to say anything. Incorporation of a repetitive skill testing in front of entire class does it for you. Running a daily test at the Rogers, or twice a day FAST like late TLG did or EL is doing now, Gabe White administering his standards both days of the class, etc puts people in their places very well. If Stoeger runs a test stage in his class in 18 sec with two Cs and a production gun at his casual pace, and best I do is 20 with two misses, an optic, and a big mag, he really doesnt need to say anything to me, let alone to a guy who runs that stage in 35 sec ...
Hi Friends: If I understand correctly, the Great Pyramid in Giza Egypt has a base that covers thirteen acres. It is about 450 feet high and weighs
Marvin Tramaine Hill is locked up on a domestic assault charge for yesterdays alleged McChicken attack in his familys Des Moines residence.
Ummm excuse me, the kid is like 1 month old - unless if you have the guns laying around in the crib (the babies, not yours) I dont see the sudden problem) Oh and what about gun safes and I guess it was OK when you had the other much older kids around. ...
Im a 19 yr old chick avec lots of time on her hands and no style. I spend a shit load of my day on Polyvore and doing other aimless things avec my aimless life ...
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REFERENCES. American Association Cereal Chemists (2000). Approved Methods of the American Association of Cereal Chemists. 10. ed. Saint Paul: Approved Methods Committee. [ Links ] Barak, S., Mudgil, D. and Khatkar, B. S. (2013). Relationship of gliadin and glutenin proteins with dough rheology, flour pasting and bread making performance of wheat varieties. LWT- Food Science and Technology Journal, 51, 211-217. http://dx.doi.org/10.1016/j.lwt.2012.09.011. [ Links ] Blechl, A. E. and Vensel, W. H. (2013). Variant high-molecular-weight glutenin subunits arising from biolistic transformation of wheat. Journal of Cereal Science, 57, 496-503. http://dx.doi.org/10.1016/j.jcs.2013.02.005. [ Links ] Branlard, G., Dardevet, M., Amiour, N. and Igrejas, G. (2003). Allelic diversity HMW and LMW glutenin subunits and Omegagliadins in French bread wheat (Triticum aestivum L.). Genetic Resources and Crop Evolution, 50, 669-679. http://dx.doi.org/10.1023/A:1025077005401. [ Links ] Costa, M. S., Scholz, M. B. S. ...
0042](1) FIGS. 1-10 illustrate a needle injection catheter 10 providing the desired features that may be constructed from the following components. As illustrated in FIGS. 1-3, a delivery tube 14 is provided. The delivery tube 14 has a first end 18, a second end 22 and a handle portion 26 adjacent the first end 18. A hypotube 30 is provided. The hypotube 30 is sized and shaped to fit slidably within the delivery tube 14, has a proximal end 34, a distal end 38 and at least three hollow needle portions 42 connected to and extending outwardly at the distal end 38. The needle portions 42, are formed of resilient material and curve outwardly from a central axis 46 of the delivery tube 14 and have distal ends 50 shaped to penetrate tissue 54 when the hypotube 30 is urged toward the second end 22 of the delivery tube 14. A first sensor 58 is provided. The first sensor 58 is located adjacent the second end 22 of the delivery tube 14 and electrically connected to a first notification device 62 located ...
We present here a transient expression system that allows the response of actin microfilaments to physiological stimuli (changes in auxin content, light) to be observed in single cells in vivo. Etiolated, intact rice seedlings are attached to glass slides, transfected biolistically with talin fused to yellow-fluorescent protein to visualize actin microfilaments, and either treated with auxin or irradiated. The talin marker labels distinct populations of actin that are differentially expressed depending on the physiological state of the coleoptile (active elongation versus ceased elongation). Whereas longitudinal transvacuolar bundles prevail in cells that have ceased to elongate, fine cortical strands are characteristic for elongating cells. The visualized actin structures remain dynamic and responsive to signals. Exogenous auxin triggers a loosening of the bundles and an extension of the cortical strands, whereas irradiation reorientates cortical strands into longitudinal arrays. These ...
The results presented point out that the Collective Field of Consciousness is a significant physical factor of the biosphere. I show that the morphogenetic field has an electromagnetic (EM) nature. EM fields are vacuum fields. Different basic forms of vacuum fields exist, and all kinds of fields, including the particle-mediated fields as well, when overlapping each other, seem to be in a direct resonant coupling, and form a complex, merged biofield. The vacuum model of consciousness presented here points to the inductive generation of consciousness, and to its self-initiating nature. Individual and collective methods, as well as the experimental possibilities of a global healing and improving the consciousness field of mankind are suggested ...
So far codon-optimized HIV-1 envelope genes have been investigated for the T cell line-adapted strain MN, which differs in several aspects from primary isolates. Envelopes of primary isolates may be more relevant for vaccine purposes. This article describes for the first time the engineering and characterization of four ""humanized"" genes encoding the secreted gp120/gp140, or the membrane-bound gp150/gp160, of a primary CCR5 tropic, clade B, clinical isolate HIV-1(BX08). The genes were built in fragments for easy cassette exchange of regions important for immunogenicity, function, and expression. The transcription and expression of the synthetic genes in mammalian cell lines were Rev independent and highly increased. Increased expression of membrane-bound gp160 induced a high cytopathic effect in U87.CD4.CCR5 cells. Gene gun and intramuscular DNA vaccination in mice induced a strong specific cytotoxic T lymphocyte response independent of the gene construct, expression level, or DNA immunization ...
Two transformation systems, via Agro bacterium tumefaciens and particle bombardment, for two varieties of chilli pepper (Capsicum annuum L.) were determined. The plasmid pCMV.CP was used for the transformation. After transformation, buds were selected on the regeneration medium containing kanamycin (50 mg/l). Based on PCR analysis, transformation via A. tumefaciens was the best approach for introducing a novel chimeric gene into chilli tissues.. ...
Women never cease to amaze me. The majority of them are still "afraid" of guns. Afraid, as in "oohhhh, they are so scary." They say things like "I dont like them around" and "theyre dangerous." I know of women who have defense-minded, pro-gun husbands and they will not let their guy talk about those nasty things in their presence. Shhh, the children will hear, or, they might actually see an evil gun! Keep them locked away. Dont tell the neighbors you have a gun in our house, or well be outcasts. And in the background you can almost hear another feeble Oprah sermon keeping women stupid, but oh, they feel good about themselves and their newfound self-esteem.. All the time, women are shocked, shocked! to hear that I have a gun, and worse, I have several guns. And then, oh no, I actually carry one with me. Thats so odd, they think. Oh shes different. Why more than one gun? She must be the aggressive type. After all, the Oprah way is to trust everyone and insist that all people have good ...
The Great Pyramid of Giza. The only one of the Seven Wonders of the Ancient World that still stands today is one of the greatest monuments ever erected on Earth. Located on the outskirts of Cairo, this pyr... ...
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Excellent Command C++, Visual Basic and Pascal.. Microsoft Office. Good ability of using the internet and searching through the web. ...
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Different national polls tend to show slightly different rates of gun ownership. The latest household gun ownership rate in the General Social Survey, in 2014, was 32 percent. The October 2015 Gallup survey showed a higher rate of 43 percent, including guns kept on property outside of the home.. But the downward trend in gun ownership remains consistent across the national polls. According to Gallup, gun ownership has fallen by about 10 percentage points since its peak in 1993. The General Social Survey shows a 20-point drop since the mid-1970s.. But gun purchases, as measured by FBI firearm background checks, are at historic highs. And data from the Bureau of Alcohol, Tobacco and Firearms shows that gun manufacturers are churning out record numbers of guns. Many gun rights advocates argue that these figures mean that the overall number of gun owners is growing: If more guns are being sold, more people must be owning guns.. But the declining rates of gun ownership across three major national ...
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We report a study of reversible adsorption of DNA-coated colloids on complementary functionalized oil droplets. We show that it is possible to control the surface coverage of oil droplets using colloidal particles by exploiting the fact that, during slow adsorption, compositional arrest takes place well before structural arrest occurs. As a consequence, we can prepare colloid-coated oil droplets with a "frozen" degree of loading but with fully ergodic colloidal dynamics on the droplets. We illustrate the equilibrium nature of the adsorbed colloidal phase by exploring the quasi-two-dimensional phase behavior of the adsorbed colloids under the influence of depletion interactions and present simulations of a simple model that illustrates the nature of the compositional arrest and the structural ergodicity. ...
In the present work we utilised some of the most discriminative molecular tools, such as RAPD, AFLP, AFRP and RAMP, to analyse the genome of independently derived transgenic plants from three elite Italian cultivars (cv. Lido, Carnaroli and Thaibonnet) and found that two methods for direct gene transfer, namely particle bombardment and intact cell electroporation (the latter being a procedure set up in this work), result in transgenic rice (Oryza sativa L.) plants that exhibit negligible genomic changes. This is in contrast with recently published results showing relevant changes in the DNA of transgenic rice plants generated through protoplasts electroporation and of transgenic poplar plants engineered through Agrobacterium tumefaciens infection. Implications of these findings are discussed in the context of selecting appropriate gene transfer methodologies to produce transgenic plants expressing genes of interest while retaining their genomic integrity and, thus, their superior agronomic ...
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In situ DNA transfer to chicken embryos by biolisticsIn situ DNA transfer to chicken embryos by biolistics

Previous reports have shown that biolistics can be used to transfer genes to chickens in ovo (Li et al., 1995; Muramatsu et al ... When transient expression is desired, in ovo lipofection, electroporation and biolistics have been used to transfect chicken ... electroporation and biolistics (Muramatsu et al., 1997). When stable gene expression and transgenic offspring are desired, stem ... embryos (Muramatsu et al., 1997). Biolistics have been used in a variety of animal tissues (Yang et al., 1990; Williams et al. ...
more infohttp://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47571999000400010

Biolistics-based gene silencing in plants using a modified par...Biolistics-based gene silencing in plants using a modified par...

Biolistics-based gene silencing in plants using a modified particle inflow gun.: RNA interference (RNAi) is one of the most ... Biolistics-based gene silencing in plants using a modified particle inflow gun.. Authors * Davies, Kevin M ...
more infohttps://www.mysciencework.com/publication/show/biolistics-based-gene-silencing-plants-using-modified-particle-inflow-gun-5268d581

An excitatory amacrine cell detects object motion and provides feature-selective input to ganglion cells in the mouse retina |...An excitatory amacrine cell detects object motion and provides feature-selective input to ganglion cells in the mouse retina |...

Using biolistics, we show that many excitatory synapses on W3-RGCs are apposed by boutons of VG3-ACs, and in whole-cell ... Biolistics. Gold particles (1.6-μm diameter, BioRad, Hercules, CA) were coated with plasmids encoding cytosolic cyan ...
more infohttps://elifesciences.org/articles/08025

Transgenesis in animal parasitic nematodes: Strongyloides spp. and Brugia spp.Transgenesis in animal parasitic nematodes: Strongyloides spp. and Brugia spp.

The method for chemically mediated transfection presented above is more versatile than biolistics, as it permits the study of ... 3.2.1. Protocol 4. Biolistics-transient transfection of isolated B. malayi embryos ...
more infohttp://wormbook.org/chapters/www_transgenesis/transgenesis.html

Sparks CA[au] - PubMed - NCBISparks CA[au] - PubMed - NCBI

Biolistics transformation of wheat.. Sparks CA, Jones HD.. Methods Mol Biol. 2009;478:71-92. doi: 10.1007/978-1-59745-379-0_4. ...
more infohttps://www.ncbi.nlm.nih.gov/pubmed?cmd=search&term=Sparks+CA%5Bau%5D&dispmax=50

METHOD OF CONFERRING MULTIPLE STRESS TOLERANCE AND EARLY FLOWERING IN PLANTS - THE GOVERNORS OF THE UNIVERSITY OF ALBERTAMETHOD OF CONFERRING MULTIPLE STRESS TOLERANCE AND EARLY FLOWERING IN PLANTS - THE GOVERNORS OF THE UNIVERSITY OF ALBERTA

Transformation methods that may be used in this regard include biolistics and Agrobacterium mediated methods. ... Res., 1988, 16:9877). Other techniques that may be used to transform dicotyledenous plant cells include biolistics (Sanford, ... biolistics); the use of naturally infective nucleic acid sequences, for example virally derived nucleic acid sequences, or ...
more infohttp://www.freepatentsonline.com/y2008/0005810.html

Glossary | Forests | Natural Resources CanadaGlossary | Forests | Natural Resources Canada

See biolistics.. Transformation par canon à particules Pathogen A microscopic organism or virus directly capable of causing ...
more infohttps://cfs.nrcan.gc.ca/terms/category/16

Cell and Molecular Laboratory | Marine Biological AssociationCell and Molecular Laboratory | Marine Biological Association

... range of microscopy and all aspects of molecular biology from PCR and electrophoresis to gene transformation and biolistics. ...
more infohttps://www.mba.ac.uk/facilities/cell-and-molecular-laboratory

Belgian Clearing-House-Mechanism : Glossary - BBelgian Clearing-House-Mechanism : Glossary - B

biolistics. in molecular biology, a method developed to inject DNA into cells by mixing the DNA with small metal particles and ...
more infohttp://bch-cbd.naturalsciences.be/belgium/glossary/glos_b.htm

BioFlorida Member Roster - BioFlorida, Inc.BioFlorida Member Roster - BioFlorida, Inc.

Boca Biolistics. Botanic Testing, Inc.. Bouchard Insurance. Brammer Bio. BridgePath Consulting. Bristol-Myers Squibb. Business ...
more infohttps://www.bioflorida.com/page/MembershipRoster

Frontiers | DNA-Free Genome Editing of Brassica oleracea and B. rapa Protoplasts Using CRISPR-Cas9 Ribonucleoprotein Complexes ...Frontiers | DNA-Free Genome Editing of Brassica oleracea and B. rapa Protoplasts Using CRISPR-Cas9 Ribonucleoprotein Complexes ...

In the other reports about the use of RNPs (Cas9 + sgRNA) in plants, biolistics was used (Svitashev et al., 2016; Liang et al ... or biolistics (Svitashev et al., 2016; Liang et al., 2017) to introduce RNPs, and some of them have succeeded in regenerating ... biolistics and PEG-mediated transformation of protoplasts. These techniques present specific concerns or have limitations in ...
more infohttps://www.frontiersin.org/articles/10.3389/fpls.2018.01594/full

Plant Breeding Correspondence Course | Plant Breeding Distance EducationPlant Breeding Correspondence Course | Plant Breeding Distance Education

Biolistics - Particle or Gene Gun. With this system, tiny heavy metal (eg. gold) micro particles are coated with the DNA to be ... The two main methods, Agrobacterium-mediated transformation and biolistics, are described below. ...
more infohttps://www.acs.edu.au/courses/plant-breeding-222.aspx

Gain-of-function Nav1.8 mutations in painful neuropathy | PNASGain-of-function Nav1.8 mutations in painful neuropathy | PNAS

2009) Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 4(8):1118-1126. ...
more infohttp://www.pnas.org/content/109/47/19444

The Domain II S4-S5 Linker in Nav1.9: A Missense Mutation Enhances Activation, Impairs Fast Inactivation, and Produces Human...The Domain II S4-S5 Linker in Nav1.9: A Missense Mutation Enhances Activation, Impairs Fast Inactivation, and Produces Human...

2009). Transfection of rat or mouse neurons by biolistics or electroporation. Nature Protocols, 4(8), 1118-1126. doi: 10.1038/ ...
more infohttps://link.springer.com/article/10.1007/s12017-015-8347-9

Photosynthetic Performance and Fertility Are Repressed in GmAOX2b Antisense Soybean | Plant PhysiologyPhotosynthetic Performance and Fertility Are Repressed in GmAOX2b Antisense Soybean | Plant Physiology

2003) Genetic transformation of soybean with biolistics. In Jackson JF, Linskens HF, eds, Molecular Methods of Plant Analysis, ... The GmAOX2b AS construct was introduced into soybean (Glycine max) via biolistics (PDS-1000/He Particle Delivery System; Bio- ...
more infohttp://www.plantphysiol.org/content/152/3/1638.full

Plus itPlus it

2009) Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 4:1118-1126, doi:10.1038/nprot.2009.90 ...
more infohttp://www.jneurosci.org/content/33/49/19250
  • Biolistics-based gene silencing in plants using a modified par. (mysciencework.com)
  • Our laboratories are exceptionally well equipped with facilities for all types of cell culturing, a wide range of microscopy and all aspects of molecular biology from PCR and electrophoresis to gene transformation and biolistics. (mba.ac.uk)
  • Scientists fire gold or tungsten microparticles from the gene gun-each microparticle coated with the gene they want to transfer into the cells using the method of biolistics. (strategicbusinessinsights.com)