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. 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)
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.
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)
A plant genus of the family POACEAE originating from the savanna of eastern Africa. It is widely grown for livestock forage.
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.
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, or their progeny, whose GENOME has been altered by GENETIC ENGINEERING.
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.
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)
Proteins found in plants (flowers, herbs, shrubs, trees, etc.). The concept does not include proteins found in vegetables for which VEGETABLE PROTEINS is available.
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)
The functional hereditary units of PLANTS.
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.
Membrane-like channels of cytoplasm connecting adjacent plant cells. Plasmodesmata connect through pores in the CELL WALL and associate with the CYTOSKELETON machinery. They are essential for intercellular transport and communication.
The process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport.
A plant family of the order Ericales, subclass Dilleniidae, class Magnoliopsida.
A plant genus of the family HAMAMELIDACEAE. The sap is a source of storax, which should not be confused with the similar named STYRAX genus.
The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments.
Components of a cell produced by various separation techniques which, though they disrupt the delicate anatomy of a cell, preserve the structure and physiology of its functioning constituents for biochemical and ultrastructural analysis. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p163)
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.
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.
Viruses which produce a mottled appearance of the leaves of plants.
Diseases of plants.
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.
A technique that labels specific sequences in whole chromosomes by in situ DNA chain elongation or PCR (polymerase chain reaction).
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.
The large scale production of pharmaceutically important and commercially valuable RECOMBINANT PROTEINS.
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.
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)
A plant genus of the family Lamiaceae. The species of Coleus should be distinguished from PLECTRANTHUS BARBATUS - which is also known as Coleus forskohlii.
Stipends or grants-in-aid granted by foundations or institutions to individuals for study.
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.
A genus of ciliate protozoa commonly used in genetic, cytological, and other research.
A species of ciliate protozoa used extensively in genetic research.
A species of ciliate protozoa used in genetic and cytological research.
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.
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.
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.
An annual legume. The SEEDS of this plant are edible and used to produce a variety of SOY FOODS.
The capacity to conceive or to induce conception. It may refer to either the male or female.
The element in plants that contains the female GAMETOPHYTES.
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)
The fertilizing element of plants that contains the male GAMETOPHYTES.
The reproductive cells of plants.

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)

Helios Gene Gun Low-Pressure System, 100-120 V from Bio-Rad,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
Transformation of Tetrahymena by microinjection of DNA was established 25 years ago. This rather labor-intensive technique has since been shelved, replaced by less time consuming and more efficient methods, electroporation and biolistics. Conjugative electroporation is the method of choice for intro …
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 ...
This study used hippocampal slice cultures in conjunction with biolistics to address the roles of PSD-95 and stargazin in controlling synaptic AMPAR localization. We demonstrate that overexpressing PSD-95 selectively enhances the AMPAR EPSC, with no change in the NMDAR EPSC. Results obtained in dissociated cultures during development suggested that PSD-95 expression required a week or more to cause changes in synaptic structure (20). However, the time course of this effect in mature neurons implies that increased levels of synaptic PSD-95 specifically recruit AMPARs to synapses, which is clearly not an effect on synaptic development. Overexpression of PSD-95 does not alter responses to exogenously applied AMPA, even though in these same cells synaptic AMPAR-mediated responses are greatly enhanced. Thus, PSD-95 appears to shift extra-synaptic AMPARs to the synapse, with no net change in total surface AMPARs. However, if synaptic AMPARs represent only a small fraction of the response to exogenous ...
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 ...
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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 ...
RNA isolation from Xenopus inner ear sensory endorgans for transcriptional profiling and molecular cloning / Casilda Trujillo-Provencio [and others] -- Synthesis of biotin-labeled RNA for gene expression measurements using oligonucleotide arrays / Ana E. Vázquez, Liping Nie, and Ebenezer N. Yamoah -- In situ hybridization approach to study mRNA expression and distribution in cochlear frozen sections / Hakim Hiel -- Lineage analysis of inner ear cells using genomic tags for clonal identification / Takunori Satoh and Donna M. Fekete -- Genetic fate-mapping approaches : new means to explore the embryonic origins of the cochlear nucleus / Jun Chul Kim and Susan M. Dymecki -- The practical use of Cre and loxP technologies in mouse auditory research / Yiling Yu and Jian Zuo -- Helios gene gun-mediated transfection of the inner ear sensory epithelium / Inna A. Belyantseva -- Electroporation-mediated gene transfer to the developing mouse inner ear / John V. Brigande [and others] -- Isolation of ...
TY - JOUR. T1 - Enhancing DNA vaccine potency by combining a strategy to prolong dendritic cell life and intracellular targeting strategies with a strategy to boost CD4+ T cell. AU - Kim, Daejin. AU - Hoory, Talia. AU - Wu, T. C.. AU - Hung, Chien Fu. PY - 2007/11/1. Y1 - 2007/11/1. N2 - Intradermal administration of DNA vaccines, using a gene gun, represents an effective means of delivering DNA directly into professional antigen-presenting cells (APCs) in the skin and thus allows the application of strategies to modify the properties of APCs to enhance DNA vaccine potency. In the current study, we hypothesized that the potency of human papillomavirus (HPV) type 16 E7 DNA vaccines employing intracellular targeting strategies combined with a strategy to prolong the life of dendritic cells (DCs) could be further enhanced by the addition of a DNA vaccine capable of generating high numbers of pan-HLA-DR reactive epitope (PADRE)-specific CD4+ T cells. We observed that the addition of PADRE DNA to E7 ...
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
WINSTON-SALEM, N.C. - Researchers at Wake Forest University School of Medicine have destroyed prostate cancer tumors in mice by injecting them with specially-coated, miniscule carbon tubes and then superheating the tubes with a brief zap of a laser.. The procedure, which used DNA-encased, multi-walled carbon nanotubes (MWCNTs) to treat human prostate cancer tumors in mice, left only a small burn on the skin that healed within days.. That we could eradicate the tumor mass and not harm the tissue is truly amazing, said principal investigator William H. Gmeiner, Ph.D., a professor of cancer biology at the School of Medicine.. Read Original Article ...
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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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
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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 ...
Figure 4: 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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Reviews of all the latest AEG and gas blow back guns in the airsoft world including carbines, sniper rifles, support weapons , sub-machine guns, pistols, and shotguns. Find out which one will give you the edge. ...
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A study published last week in the journal Science links a surge in gun sales post-Sandy Hook to a rise in accidental shooting deaths.
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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 ...
APRIL 4--After discovering this classic mug shot, we doubted that another arrestee would ever be photographed in a similar state.
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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.
Physical methods like electroporation, biolistics ("gene gun"), sonoporation that uses cavitation of gas bubbles produced by ... or biolistics. Particles of gold or tungsten are coated with DNA and then shot into young plant cells or plant embryos. Some ...
Other methods include biolistics, where particles of gold or tungsten are coated with DNA and then shot into young plant cells, ...
Gene guns (also known as biolistics) "shoot" (direct high energy particles or radiations against[49]) target genes into plant ...
Genetic material can also be inserted into a tomato cell's chloroplast and chromoplast plastomes using biolistics. Tomatoes ...
The approaches taken by governments to assess and manage the risks associated with the use of genetic engineering technology and the development and release of GMOs vary from country to country, with some of the most marked differences occurring between the United States and Europe. The U.S. regulatory policy is governed by the Coordinated Framework for Regulation of Biotechnology[48] The policy has three tenets: "(1) U.S. policy would focus on the product of genetic modification (GM) techniques, not the process itself, (2) only regulation grounded in verifiable scientific risks would be tolerated, and (3) GM products are on a continuum with existing products and, therefore, existing statutes are sufficient to review the products."[49] European Union by contrast enacted regulatory laws in 2003 that provided possibly the most stringent GMO regulations in the world.[5] All GMOs, along with irradiated food, are considered "new food" and subject to extensive, case-by-case, science-based food ...
... tumefaciens causes crown-gall disease in plants. The disease is characterised by a tumour-like growth or gall on the infected plant, often at the junction between the root and the shoot. Tumors are incited by the conjugative transfer of a DNA segment (T-DNA) from the bacterial tumour-inducing (Ti) plasmid. The closely related species, Agrobacterium rhizogenes, induces root tumors, and carries the distinct Ri (root-inducing) plasmid. Although the taxonomy of Agrobacterium is currently under revision it can be generalised that 3 biovars exist within the genus, Agrobacterium tumefaciens, Agrobacterium rhizogenes, and Agrobacterium vitis. Strains within Agrobacterium tumefaciens and Agrobacterium rhizogenes are known to be able to harbour either a Ti or Ri-plasmid, whilst strains of Agrobacterium vitis, generally restricted to grapevines, can harbour a Ti-plasmid. Non-Agrobacterium strains have been isolated from environmental samples which harbour a Ri-plasmid whilst laboratory ...
In 1978 Bayer purchased Miles Laboratories and its subsidiaries Miles Canada and Cutter Laboratories, acquiring along with them a variety of product lines including Alka-Seltzer, Flintstones vitamins and One-A-Day vitamins, and Cutter insect repellent.[53] Along with the purchase of Cutter, Bayer acquired Cutter's Factor VIII business. Factor VIII, a clotting agent used to treat hemophilia, was produced, at the time, by processing donated blood. In the early days of the AIDS epidemic, people with hemophilia were found to have higher rates of AIDS, and by 1983 the CDC had identified contaminated blood products as a source of infection. According to the New York Times, this was "one of the worst drug-related medical disasters in history". Companies, including Bayer, developed new ways to treat donated blood with heat to decontaminate it, and these new products were introduced early in 1984. In 1997 Bayer and the other three makers of such blood products agreed to pay $660 million to settle cases ...
Much debate surrounds the topic of human enhancement and the means used to achieve one's enhancement goals.[19] In some circles the expression "human enhancement" is roughly synonymous with human genetic engineering,[20][21] but most often it is referred to the general application of the convergence of nanotechnology, biotechnology, information technology and cognitive science (NBIC) to improve human performance.[22] Since the 1990s, several academics (such as some of the fellows of the Institute for Ethics and Emerging Technologies[23]) have risen to become advocates of the case for human enhancement while other academics (such as the members of President Bush's Council on Bioethics[24]) have become outspoken critics.[25] Advocacy of the case for human enhancement is increasingly becoming synonymous with "transhumanism", a controversial ideology and movement which has emerged to support the recognition and protection of the right of citizens to either maintain or modify their own minds and ...
The term eugenics and its modern field of study were first formulated by Francis Galton in 1883,[69] drawing on the recent work of his half-cousin Charles Darwin.[70][71] Galton published his observations and conclusions in his book Inquiries into Human Faculty and Its Development. The origins of the concept began with certain interpretations of Mendelian inheritance and the theories of August Weismann.[72] The word eugenics is derived from the Greek word eu ("good" or "well") and the suffix -genēs ("born"), and was coined by Galton in 1883 to replace the word "stirpiculture", which he had used previously but which had come to be mocked due to its perceived sexual overtones.[73] Galton defined eugenics as "the study of all agencies under human control which can improve or impair the racial quality of future generations".[74] Historically, the term eugenics has referred to everything from prenatal care for mothers to forced sterilization and euthanasia.[75] To population geneticists, the term ...
There is variability in the whole procedure depending largely on the strain from which the stem cells have been derived. Generally cells derived from strain 129 are used. This specific strain is not suitable for many experiments (e.g., behavioural), so it is very common to backcross the offspring to other strains. Some genomic loci have been proven very difficult to knock out. Reasons might be the presence of repetitive sequences, extensive DNA methylation, or heterochromatin. The confounding presence of neighbouring 129 genes on the knockout segment of genetic material has been dubbed the "flanking-gene effect".[6] Methods and guidelines to deal with this problem have been proposed.[7][8] Another limitation is that conventional (i.e. non-conditional) knockout mice develop in the absence of the gene being investigated. At times, loss of activity during development may mask the role of the gene in the adult state, especially if the gene is involved in numerous processes spanning development. ...
In addition to being practiced in a number of countries, eugenics was internationally organized through the International Federation of Eugenics Organizations.[28] Its scientific aspects were carried on through research bodies such as the Kaiser Wilhelm Institute of Anthropology, Human Heredity, and Eugenics,[29] the Cold Spring Harbour Carnegie Institution for Experimental Evolution,[30] and the Eugenics Record Office.[31] Politically, the movement advocated measures such as sterilization laws.[32] In its moral dimension, eugenics rejected the doctrine that all human beings are born equal and redefined moral worth purely in terms of genetic fitness.[33] Its racist elements included pursuit of a pure "Nordic race" or "Aryan" genetic pool and the eventual elimination of "unfit" races.[34][35] Many leading British politicians subscribed to the theories of eugenics. Winston Churchill supported the British Eugenics Society and was an honorary vice president for the organization. Churchill believed ...
A gene knockout (abbreviation: KO) is a genetic technique in which one of an organism's genes is made inoperative ("knocked out" of the organism). However, KO can also refer to the gene that is knocked out or the organism that carries the gene knockout. Knockout organisms or simply knockouts are used to study gene function, usually by investigating the effect of gene loss. Researchers draw inferences from the difference between the knockout organism and normal individuals. The KO technique is essentially the opposite of a gene knock-in. Knocking out two genes simultaneously in an organism is known as a double knockout (DKO). Similarly the terms triple knockout (TKO) and quadruple knockouts (QKO) are used to describe three or four knocked out genes, respectively. However, one needs to distinguish between heterozygous and homozygous KOs. In the former, only one of two gene copies (alleles) is knocked out, in the latter both are knocked out. ...
Until the 1990s, Europe's regulation was less strict than in the United States, one turning point being cited as the export of the United States' first GM-containing soy harvest in 1996. The GM soy made up about 2% of the total harvest at the time, and Eurocommerce and European food retailers required that it be separated.[2] In 1998, the use of MON810, a Bt expressing maize conferring resistance to the European corn borer, was approved for commercial cultivation in Europe. Shortly thereafter, the EU enacted a de facto moratorium on new approvals of GMOs pending new regulatory laws passed in 2003. Those new laws provided the European Union (EU) with possibly the most stringent GMO regulations in the world.[1] All GMOs, along with irradiated food, are considered "new food" and subject to extensive, case-by-case, science based food evaluation by the European Food Safety Authority (EFSA). The criteria for authorization fall in four broad categories: "safety", "freedom of choice", "labelling" and ...
In 1974 Rudolf Jaenisch created a transgenic mouse by introducing foreign DNA into its embryo, making it the world's first transgenic animal.[14][15] However it took another eight years before transgenic mice were developed that passed the transgene to their offspring.[16][17] Genetically modified mice were created in 1984 that carried cloned oncogenes, predisposing them to developing cancer.[18] Mice with genes knocked out (knockout mouse) were created in 1989. The first transgenic livestock were produced in 1985[19] and the first animal to synthesise transgenic proteins in their milk were mice,[20] engineered to produce human tissue plasminogen activator in 1987.[21]. In 1983 the first genetically engineered plant was developed by Michael W. Bevan, Richard B. Flavell and Mary-Dell Chilton. They infected tobacco with Agrobacterium transformed with an antibiotic resistance gene and through tissue culture techniques were able to grow a new plant containing the resistance gene.[22] The gene gun ...
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. ...
Transformations using biolistics are simple and efficient, making it the method of choice. This is especially important for low ... Transformation of DNA into Tetrahymena is performed using microinjection, electroporation, or biolistics. Transformations ... whereas electroporation and biolistics rely on the precise timing of the transformation during the cell cycle or during ...
This technique is often simply referred to as bioballistics or biolistics.. This device is able to transform almost any type of ...
Transgenic trees currently are being produced by Agrobacterium-mediated transformation and biolistics. Since trees are ...
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 ...
Biolistics. A rat myosin Va cDNA clone containing the coil-coil and globular tail, but lacking the IQ repeats and motor domain ...
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 ...
New genetic information can be inserted with biolistics. An example of transgenics is the rainbow papaya, which is modified ...
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 OBrien presents...Gene Gun Barrels for more information about biolistics. ...
Biolistics transformation of wheat.. Sparks CA, Jones HD.. Methods Mol Biol. 2009;478:71-92. doi: 10.1007/978-1-59745-379-0_4. ...
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 ...
Biolistics * Biological Transport * Green Fluorescent Proteins * Luminescent Proteins / genetics * Plant Proteins / metabolism ...
Biolistics * Cell Proliferation * Complement C3d / immunology * DNA / administration & dosage* * Female * Hemagglutinin ...
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 ...
See biolistics.. Transformation par canon à particules Pathogen A microscopic organism or virus directly capable of causing ...
Other methods include biolistics, where particles of gold or tungsten are coated with DNA and then shot into young plant cells, ...
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 ...
2009) Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 4(8):1118-1126. ...
2009). Transfection of rat or mouse neurons by biolistics or electroporation. Nature Protocols, 4(8), 1118-1126. doi: 10.1038/ ...
Two days later, slices were transfected via particle-based biolistics. Slices were imaged at 4 DIV via confocal microscope. ...
BIOLISTICS. *TALENS AND ZFNS*Table MICE MODEL MARKET SIZE FOR OTHER TECHNOLOGIES, BY COUNTRY, 2014-2021 (USD THOUSAND) ...
... range of microscopy and all aspects of molecular biology from PCR and electrophoresis to gene transformation and biolistics. ...
biolistics. in molecular biology, a method developed to inject DNA into cells by mixing the DNA with small metal particles and ...
... used to transform wheat using biolistics; shown are the probe region for Southern blot and primer sequences FgPf1 and M13RevP ...
Construction of a candidate vaccine against avian influenza, strain H5N1 via biolistics mediation. In: The 21st Intervarsity ...
2009) Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 4:1118-1126, doi:10.1038/nprot.2009.90 ...
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. ...
Boca Biolistics. Botanic Testing, Inc.. Brammer Bio. Brand Institute, Inc.. BridgePath Consulting. Bristol-Myers Squibb. ...
Figure 4a depicts the construct utilized for targeted engineering of the P. renovo chloroplast via biolistics. The native 16S ... 3b). Transgene integration was also achieved via biolistics, however we observed approximately an order of magnitude lower ...
Animals, Biolistics, Caenorhabditis elegans, Culture Media, DNA, Eggs, Gold, Transformation, Genetic, Journal Article ...
  • Other techniques used successfully in poultry include cell transfection with retroviral vectors, lipofection in ovo , electroporation and biolistics (Muramatsu et al . (scielo.br)
  • Alternative techniques like chemical sensitivation of cells, electroporation and biolistics can also be used. (wisegeek.org)
  • These include methods involving spheroplast generation, electroporation, and biolistics with DNA coated microprojectiles. (asmscience.org)
  • 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)
  • Biolistics is an efficient method for transformation of 'Chancellor' and should be applicable to other important grape cultivars. (springer.com)
  • Gene editing reagents are often still delivered to plant callus cells in culture using decades-old technology, such as Agrobacterium- and biolistics (transformation via ballistic DNA delivery)-based approaches. (genengnews.com)
  • Information and tips on electroporation, viral transfection vector, lipofection, and biolistics transformation for planning and selecting the appropriate transfection method. (bio-rad.com)
  • The most common delivery method is biolistics based transformation but due to the high frequency of gene silencing linked with high copy transgenes and low edit rate in wheat, a large population of transgenic plants are needed for recovery of desired mutations. (isaaa.org)
  • In many plants, including rice, Agrobacterium -mediated transformation is the most practical means of transformation because this biotic transformation system can deliver longer and more intact DNA payloads with less incorporation of fragmented DNA compared with physical transformation systems such as polyethylene glycol, electroporation, or biolistics. (plantphysiol.org)
  • The method used mostly is particle bombardment (biolistics). (brokenfadercartel.com)
  • The particle-mediated DNA transfer, also refers to gene gun, biolistic particle delivery, biolistics or particle bombardment, is one the commonly used methods contribute to manipulating the gene of organisms. (creative-biolabs.com)
  • We built Boca Biolistics' internal reference lab to meet the needs of TODAY'S clinicians and tomorrow's health care research. (bocabio.com)
  • That means every health care research dollar goes further when you work with the team at Boca Biolistics. (bocabio.com)
  • Boca Biolistics reference lab is constantly collecting new data points on our library of samples. (bocabio.com)
  • Get in touch with Boca Biolistics to discuss our reference lab solutions today. (bocabio.com)
  • 2009). Transfection of rat or mouse neurons by biolistics or electroporation. (springer.com)
  • We use techniques such as biolistics transfection to see if the overexpression of genes that might be beneficial for retinal neurons can result in cell rescue (Insert figure here). (cardiff.ac.uk)
  • Some plant species are not susceptible to Agrobacterium DNA integration, and in that case, we could use an alternative method for transforming a plant such as biolistics. (biotechlearn.org.nz)
  • Biolistics-based gene silencing in plants using a modified par. (mysciencework.com)
  • The construct containing a YFP gene was used to transfect embryos via biolistics to test whether YFP and GLuc are expressed. (usf.edu)
  • The plasmids pULGU1 and pEmuGN were introduced by biolistics in embryogenic cell suspensions of pearl millet, Pennisetum glaucum. (academicjournals.org)
  • Examination of randomly selected transformants by biolistics showed that 15 to 40% were stable, depending on the recipient auxotroph, with integrative events identified in all stable transformants by DNA analysis. (asm.org)
  • In biolistics, the process starts by mixing a DNA construct with particles of a heavy metal, usually tungsten or gold. (goldbio.com)
  • Biolistics is like a small gun that shoots tiny gold particles that are coated in DNA into the plant, and if you shoot enough of these particles into the leaf, for example, of a plant, some of it will get integrated and you get transformed cells. (biotechlearn.org.nz)
  • In addition to inventing the Nanopatch - and driving it forward towards product - Mark also contributed to the biolistics technology while serving as a Lecturer at Oxford University. (weforum.org)
  • The biolistics technology was spun out to PowerJect (sold to Chiron for $1 billion (in 2003)) and then, as PowderMed to Pfizer for $400 million (in 2006). (weforum.org)
  • While at the University of Oxford, I was an inventor of the biolistics technology, commercialised with PowderJect (sold to Chiron Vaccines for $1 Billion in 2003), and then PowderMed, purchased by Pfizer for $400 million in 2006. (mysecuritymarketplace.com)