There is a hygromycin marker on the pGIPZ, pTRIPZ, and pLemiR vectors. It was present on the parent plasmid that they were constructed from but its function has never been tested in house. The hygromycin in the TRIPZ vector was specifically inactivated so it will not work. In theory you could use the hygromycin marker for selection of cells transfected with the pGIPZ or pLemiR plasmid but the hygromycin takes longer to have the desired effect than puromycin. You would not be able to use hygromycin for selection of transduced cells because only what is between the LTRs of the vectors would be packaged and the hygromycin marker lies outside of the LTRs ...

0159] Resistance genes for glyphosate (resistance conferred by mutant 5-enolpyruvl-3 phosphikimate synthase (EPSP) and aroA genes, respectively), and hygromycin B phosphotransferase, and to other phosphono compounds such as glufosinate (phosphinothricin acetyl transferase (PAT) and Streptomyces hygroscopicus phosphinothricin-acetyl transferase (bar) genes) may also be used. See, for example, U.S. Pat. No. 4,940,835 to Shah, et al., which discloses the nucleotide sequence of a form of EPSPS which can confer glyphosate resistance. A DNA molecule encoding a mutant aroA gene can be obtained under ATCC accession number 39256, and the nucleotide sequence of the mutant gene is disclosed in U.S. Pat. No. 4,769,061 to Comai. A hygromycin B phosphotransferase gene from E. coli that confers resistance to glyphosate in tobacco callus and plants is described in Penaloza-Vazquez et al. (Plant Cell Reports, 14:482-487, 1995). European patent application No. 0 333 033 to Kumada et al., and U.S. Pat. No. ...

Hygromycin B solution is a selection antibiotic for mammalian and bacterial cells. It is sterile, cell culture tested, and available as a solution. CAS 31282-04-9

In experiments with albino rats it was found that after administration of phytobacteriomycin, trichotecin, hygromycin B or levoristatin into the stomach in doses of 1/20 of LD50 activity of the microsomal enzymes of the liver cells significantly changed and the changes persisted within at least 2 weeks. The above antibiotics induced similar changes in the lysosome enzyme, i.e. acid phosphatase, providing an increase in its activity. Changes in the activity of succinate dehydrogenase (mytochondria indicator enzyme), glucose-6-phosphatase (ribosome indicator enzyme) and aspartate aminotransferase (cytoplasm indicator enzyme) were different for each antibiotic. It is concluded that the above antibiotics were capable of impairing on intoxication the enzymatic function of various cell microstructures, though the levels of the change direction may be different.

Comparison of yeast cell growth upon hygromycin B treatment.Serial dilutions of the strains were grown on YPD plates containing 0 (control), 30 and 50 µg/ml hy

Clontech offers a variety of antibiotics for use in cell culture and bacterial selection. Antibiotic selection drugs include puromycin, hygromycin, G418 geneticin, neomycin and doxycycline.

Clontech offers a variety of antibiotics for use in cell culture and bacterial selection. Antibiotic selection drugs include puromycin, hygromycin, G418 geneticin, neomycin and doxycycline.

Curiously, instead of conferring tolerance to moderate Na+ stress or high K+, CHX20 consistently caused mutant KTA40-2 (Δena1-4 Δnha1 Δnhx1 Δkha1) to be more sensitive to salt. In another salt-sensitive yeast mutant, AXT3 (Δena1-4 Δnha1 Δnhx1), expression of AtCHX20 also resulted in increased sensitivity to moderate Na+ stress and high K+, although AtNHX1 or AtNHX2 conferred moderate tolerance to Na+ stress (data not shown) as shown before (Yokoi et al., 2002). Furthermore, CHX20 was unable to confer hygromycin B tolerance. Thus, AtCHX20 is functionally distinct from the vacuolar AtNHX1 that sequesters excess Na+ or K+ into vacuoles and confers tolerance to high Na+ or K+ and to hygromycin B (Pardo et al., 2006).. Instead, CHX20 function appears to be important particularly when K+ is depleted and when the external pH is slightly alkaline. This is shown by improved growth of KTA40-2 expressing CHX20 at pH 7.5 and when [K+]ext was low (between 0.4 and 3 mm). Yeast growth and budding ...

|p|Application: Recommended for use as a selection agent at 100-800 μg/mL. Biochem/physiol Actions: Mode of Action: Blocks polypeptide synthesis and inhibits elongation. For use in the selection and maintenance of prokaryotic and eukaryotic cells. Preparation Note: Purified by ion exchange chromatography|/p|

Lab ID pUMa194 (short hyg promoter version), published as pBS-hhn, this plasmid was not developed in our lab (this plasmid is easier to use than the original, larger plasmid pMF1-h Brachmann et al., 2004 ...

ID PHPHP1 preliminary; circular DNA; SYN; 3818 BP. XX AC IG0168; K01193; X01385; XX DT 02-NOV-1992 (Rel. 7, Created) DT 01-JUL-1995 (Rel. 12, Last updated, Version 1) XX DE E. coli phagemid vector pHph+1 - complete. XX KW cloning vector. XX OS Cloning vector OC Artificial sequences; Cloning vehicles. XX RN [1] RC pHph+1 RA Muller A., Piepersberg W.; RT ; RL Submitted (08-AUG-1989) by: RL Muller A., Inst. fur Biochemie, Darmstadt, Germany. XX RN [2] RC pHph-1, pHph+1 from pUC9 & hygromycin resistance gene RA Muller A., Piepersberg W.; RT ; RL Unpublished (1984). XX CC old Boehringer vector. CC NM (pHph+1) CC CM (yes) CC NA (ds-DNA) CC TP (circular) CC ST () CC TY (phagemid) CC SP () CC HO (E.coli K-12) CC CP () CC FN (promoter analysis) CC SE () CC PA (pUC8)(pJHPH) CC BR (pHph0)(pHph-1) CC OF () CC OR () XX FH Key Location/Qualifiers FH FT misc_feature 0..0 FT /note=1. E. coli 1142bp, hyg gene FT -, pJHph FT 1. pUC12 remove EcoRI-BamHI 18bp 231..249, MCS/2662bp FT 2. pJHph 1142bp 220..1362, hyg ...

pORF39-Hph from InvivoGen,Hygromycin resistance gene (hph), provided in a pORF plasmid. pORF is an expression plasmid selectable with Ampicillin.,biological,biology supply,biology supplies,biology product

This line was derived from the human embryonic kidney line, 293 in 2001. 293 cells were transfected with the plasmid pVgRXR bearing a Zeocin-resistance selectable marker to obtain 293VgRXR cells. This population was subsequently transfected with the plasmid pEKORF6 containing Ad5 E4 ORF 6 that encodes E4 34K. pEKORF6 is a derivative of the plasmid pIndHydro that contains a hygromycin resistance gene. The vectors contain SV40 viral DNA sequences. The 2V6.11 cell line was originally selected in hygromycin-containing medium and cloned by single-colony isolation with a cloning cylinder. The 2V6.11 cells inducibly express the human adenovirus E4, 34kDa protein (E4 ORF6). 2V6.11 cells exhibit little or no constitutive E4 biologic activity. Induction by ponasterone A, an ecdysone analog, however, results in E4 biologic activity and levels of E4 34kDa protein that are detectable by immunoblotting. These cells can be used as a tool to study the biology of the adenoviral E4 oncoprotein.

Dear SVemuri12, I suspect that you are using too high an annealing temperature. The calculated Tm is just an estimate. Also, my rule of thumb for annealing temperatures is Tm - 5°C (cant remember where I picked up that bit of information). Try dropping the annealing to 55°C, or even lower. Also make sure you are using a sufficient number of cycles, that the PCR recipe is OK, PCR machine is working properly, pipettors are OK, DNA template is good etc. The conditions described for these primers by Chen et al (2006) seem a bit more stringent than is normally needed. Use a standard three temperature PCR (95°C, 5 mins; 94°C 30 secs, 55°C 30 secs, 72°C 1 min for 35 cycles; end with 72°C for 10 min). Good luck, Vince Vincent Mulholland Molecular Biology Unit Manager, Diagnostics & Molecular Biology Section, Scottish Agricultural Science Agency Hello, To those who have worked with hygromycin primers for genomic DNA: I am trying to do PCR amplification of my genomic DNA (WT and mutant lines). So ...

The inositol portion was synthesised using the key TA step which had an superb yield of 74% on using (only) 1 mol% catalyst loading, giving the desired diastereomer exclusively. This step has been improved upon the previously reported result (61% yield at 4 mol% catalyst loading). The attacment of the inositol portion to the B+C part was achieved using standard coupling reagents and on deprotection yielded Hygromycin A in 17 linear steps and 10% overall yield a great improvement over the previous synthesis ...

The effect of apramycin treatment on transfer and selection of an Escherichia coli strain (E. coli 912) in the intestine of pigs was analyzed through an in vivo experiment. The strain was sequenced and assigned to the sequence type ST101 and serotype O11. It carried resistance genes to apramycin/gentamicin, sulphonamide, tetracycline, hygromycin B, β-lactams and streptomycin [aac(3)-IV, sul2, tet(X), aph(4), bla TEM-1 and strA/B], with all but tet(X) located on the same conjugative plasmid. Nineteen pigs were randomly allocated into two inoculation groups, one treated with apramycin (pen 2) and one non-treated (pen 3), along with a non-inoculated control group (pen 1 ...

Episomal mammalian vector with a CMV promoter and a hygromycin resistance marker, for high-level expression in primate and canine cells.

We are trying to establish two stable cell lines using G418 resistance and hygromycin resistance. After determining the concentration of each drug needed to kill the cells, we proceeded with transfections. Some of the control cells (approx. 30%), those transfected with no DNA, did not die. Has anyone encountered this problem before? If so, what actions did you take? Could you still establish a stable cell line? Thanks. http://biowww.net/mynews/tree.php?group_name=bionet_immunology&begin=0 ...

Mycoplasma coexpression expression vector with the Phsp promoter and C-termial c-myc tag; newer version of pMYC4-H suitable for universal cloning by PacI and SbfI sites; hygromycin resistance in bacteria and mycobacteria; restriction enzyme cloning (PacI, SbfI ...

HEK 293T/17 cells were transformed with adenovirus E1a carrying a temperature sensitive T antigen co-selected with neomycin. Transformation was brought about by the insertion of approximately 4.5 kilobases of viral genome into human chromosome 19. Gag-pol was introduced with hygromycin as the co-selectable marker and the envelope proteins were introduced with diptheria resistance as the co-selectable marker.

I am trying to determine the best dose of hygromycin that i can use to select my transfected colonies. (working with chinese hamster cell lines). I have tried several concentrations (from 50-250 microgram per ml) on my host cells and changing the media plus antibiotic every 1-2 days. Each time the media has turned yellow and lots of dead cells but still surviving colonies can be seen. How could we know if the cell death is because of the overgrowth of the culture and consequent bad condition or because of the antibiotic?. ...

The NYSE volume today was abysmal. According to BBG data, this was the lowest volume day in over a decade and even compared to other July 1st (or holiday weeks) this was the lowest volume print. Average trade size for the S&P 500 e-mini futures was also very small - almost the lowest of the year as low volumes and the narrowest high-to-low range for ES in over two months still managed to hold on to small gains for the day. In the face of this relative exuberance, Treasury yields dumped down 5 to 6 bps across the board remaining the most cognitively dissonant of risk assets on the day. HYG underperformed (as HY and IG credit indices were very quiet and reracked along with ES for most of the day). HYG did end Friday notably rich to intrinsics so this makes some sense but is unusual for a positive close in ES (as we note that 16 of the 24 times in the last year that HYG has closed red and SPY closed green, SPY has gone on to lose more in the next few days). EURUSD lost quite a bit of ground (again

Van Der Straten, A., Johansen, H., Sweet, R., & Rosenberg, M. (1987). Efficient expression of foreign genes in cultured drosophila melanogaster cells using hygromycin B selection. In Y. Kuroda, E. Kurstak, & K. Maramorosch (Eds.), Invertebrate and Fish Tissue Culture: Proceedings of the Seventh International Conference on Invertebrate and Fish Tissue Culture, Japan (pp. 131-134). New York, NY: Springer Verlag ...

Three transformation systems have been reported for the rice blast fungus Magnaporthe grisea (Parsons et al. 1987 Proc. Natl. Acad. Sci. USA 84:4161-4165; Daboussi et al. 1989 Curr. Genet. 15:453-456; Leung et al. 1990 Curr. Genet. 17:409-411). Among these three selection systems, only hygromycin B resistance provides a dominant selection that can be used for any wild type strain. A second dominant selection marker is needed to transform strains that are already hygromycin B resistant.. Bialophos produced by Streptomyces hygroscopicus is a tripeptide consisting of two L- alanine residues and an analogue of glutamic acid called phosphinothricin (PPT). Upon cleavage of bialophos, PPT is released and acts as an inhibitor of glutamine synthesis in plants, and hence functions as a potent herbicide. A gene (bar) coding for PPT acetyltransferase has been isolated from S. hygroscopicus and is widely used as a selective marker for the transformation of higher plants (DeBlock et al. 1987 EMBO J. ...

Transformed Arabidopsis thaliana plants have been produced by a modified leaf disk transformation-regeneration method. Leaf pieces from sterilely grown plants were precultured for 2 days and inoculated with an Agrobacterium tumefaciens strain containing an avirulent Ti (tumor-inducing) plasmid with a chimeric gene encoding hygromycin resistance. After cocultivation for 2 days, the leaf pieces were placed on a medium that selects for hygromycin resistance. Shoots regenerated within 3 months and were excised, rooted, and transferred to soil. Transformation was confirmed by opine production, hygromycin resistance, and DNA blot hybridization of both primary transformants and progeny. This process for producing transgenic Arabidopsis plants should enhance the usefulness of the species for experimental biology. |P /|

Neuss, N. and Koch, K. F. and Molloy, B. B. et al. (1970) Structure of hygromycin B, an antibiotic from Streptomyces hygroscopicus; The use of CMR. spectra in structure determination, I. Helvetica Chimica Acta, 53 (8). pp. 2314-2319. ISSN 0018-019X. https://resolver.caltech.edu/CaltechAUTHORS:20151023-133453240 Dorman, Douglas E. and Angyal, S. J. and Roberts, John D. (1970) Nuclear Magnetic Resonance Spectroscopy. Carbon-13 Spectra of Some Inositols and Their O-Methylated Derivatives. Journal of the American Chemical Society, 92 (5). pp. 1351-1354. ISSN 0002-7863. https://resolver.caltech.edu/CaltechAUTHORS:20151013-111511440 Dorman, Douglas E. and Roberts, John D. (1970) Nuclear Magnetic Resonance Spectroscopy. Carbon-13 Spectra of Some Pentose and Hexose Aldopyranoses. Journal of the American Chemical Society, 92 (5). pp. 1355-1361. ISSN 0002-7863. https://resolver.caltech.edu/CaltechAUTHORS:20151013-111511687 Dorman, Douglas E. and Roberts, John D. (1970) Nuclear magnetic resonance ...

EN] Potassium homeostasis is crucial for living cells. In the yeast Saccharomyces cerevisiae, the uptake of potassium is driven by the electrochemical gradient generated by the Pma1 H +-ATPase, and this process represents a major consumer of the gradient. We considered that any mutation resulting in an alteration of the electrochemical gradient could give rise to anomalous sensitivity to any cationic drug independently of its toxicity mechanism. Here, we describe a genomewide screen for mutants that present altered tolerance to hygromycin B, spermine, and tetramethylammonium. Two hundred twenty-six mutant strains displayed altered tolerance to all three drugs (202 hypersensitive and 24 hypertolerant), and more than 50% presented a strong or moderate growth defect at a limiting potassium concentration (1 mM). Functional groups such as protein kinases and phosphatases, intracellular trafficking, transcription, or cell cycle and DNA processing were enriched. Essentially, our screen has identified a ...

1HNX: The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit.

2016-6-15 · Chloramphenicol, USP Grade C-105-5 5 g 11 C-105-25 25 g 21 C-105-100 100 g 74 G-418 Sulfate G-418-5 5 g 129 G-418-10 10 g 216 G-418-25 25 g 318 Hygromycin B** H-270-1 1 g 64 H-270-2.5 2.5 g 151 H-270-5 5 g 265 H-270-10 10 g 477 Kanamycin Monosulfate K-120-5 5 g 21 K-120-10 10 g 41. Get Price ...

Drugs. 2,3-Dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide (NBQX) from Ascent Scientific (Weston-SuperMare, UK). Dihydro-β-erythroidine hydrobromide (DHβE), methyllycaconitine (MLA), and PNU-120596 were purchased from Tocris Bioscience (Bristol, UK). SB-206553, picrotoxin, atropine, (-)nicotine, and acetylcholine were purchased from Sigma Chemical (Poole, Dorset, UK).. Recombinant and Native Cell Lines. GH4C1 cells stably transfected with pCEP4/rat α7 nAChR (α7-nAChR-GH4C1) were used in this study and maintained in poly-d-lysine-coated flasks in F10 medium supplemented with 15% horse serum and 2.5% fetal bovine serum, 1% penicillin-streptomycin, and 200 mg/ml hygromycin B at 37°C in a humidified 5% CO2 incubator. The 5-HT3A receptor cDNA was cloned from human brain RNA, and the rat α7 nAChR was cloned from PC12 cells. Human SHSY5Y cells and TE671 endogenously expressing α3- and α1-containing receptors, respectively, were used.. Measurement of Intracellular Ca2+Using the ...

Targeted gene and genome manipulation via homologous recombination in bacteria relies on the use of marker genes. In marker-less gene replacement, a negative and a positive selection marker are employed, in a process that normally involves two transformation steps with two DNA suicide vectors. In this work, we developed an alternative strategy for marker-less gene replacement in Synechocystis, based on the use of a single plasmid and a single transformation step. The technique was tested by inserting the luxAB operon from Vibrio fischeri into the neutral receptor site slr0168. To this end the wild-type strain was transformed with the vector pDSlux and the transformation efficiency with this integration vector was in the range of 10-5, while the average frequency of the second recombination event was 10-6 (see Table 1). This ten-fold difference could be due to the fact that the first recombination event involves a double crossover between the genomic DNA and the plasmid vector, whereas the second ...

Pharmaceutical Frontier Research Laboratory, Central Pharmaceutical Research Institute, Japan Tobacco Inc., 13-2, Fukuura 1-chome, Kanazawa-ku, Yokohama, Kanagawa, Japan. We developed a simple and rapid method of constructing knockout vectors using inverse-PCR (IPCR). The method consists of 3 steps: 1) digestion of a target bacterial artificial chromosome (BAC) with several restriction enzymes (six-base cutters) followed by self-ligation, 2) IPCR using circular DNAs as templates and two primers which are oriented in the reverse direction and 3) cloning into a vector containing a positive selection marker, which results in a typical replacement knockout vector. We successfully targeted 3 mouse genes including the HPRT gene by using this method. Compared with the conventional method, this method has several advantages. First, it is relatively quick. While the conventional method requires several months for completion of a targeting vector, this novel method requires no more than 3 weeks. Second, ...

0044] RT-PCR. RNA was purified with Tri-reagent (Sigma, St. Louis, Mo.). cDNA was synthesized from 1 μg of RNA by with PROTOSCRIPT First Strand cDNA Synthesis Kit (NEB, Ipswich, Mass.). PCR was performed with 1 μl of cDNA with forward and reverse primers specific for GAPDH (5-TGC ATC CTG CAC CAC CAA CT-3, SEQ ID NO:1; and 5-TGC CTG CTT CAC CAC CTT C-3, SEQ ID NO:2), hygromycin phosphotransferase (5-CAT GGC GTG ATT TCA TAT GCG CGA-3, SEQ ID NO:3; and 5-TCC AGA AGA AGA TGT TGG CGA CCT-3, SEQ ID NO:4), puromycin acetyltransferase (5-ACC GAG CTG CAA GAA CTC TTC CTC-3, SEQ ID NO:5; and 5-AGG AGG CCT TCC ATC TGT TGC T-3, SEQ ID NO:6), Pdpn (5-ACC AAC ACA GAC GAC CAA GAC ACT-3, SEQ ID NO:7; and 5-AAG CAT CCA CTG TGC CTT CAG TTC-3, SEQ ID NO:8), Fhl1 (5-GAG AAG TTC GAC TGT CAC TAC TGC-3, SEQ ID NO:9; and 5-CTG ATC CTG GTA AGT GAT TCC TCC-3, SEQ ID NO:10), 4930408021Rik (5-TGT TCT CAG AGC CCA GCA TCA CTT-3, SEQ ID NO:11; and 5-ACA TCC TCT CAG CTG GTT CCT TCA-3, SEQ ID NO:12), ...

Coloured scanning electron micrograph (SEM) of Streptomyces hygroscopicus, Gram-positive, aerobic, filamentous, biofilm-forming, rod prokaryote (bacterium). Streptomyces sp. belong to the Actinomycetes group and are bacteria that share many characteristics with the fungi. They grow usually as filaments (chains of cells) and often branch to form a network of filaments (mycelium) in the soil. These soil bacteria are responsible for the musty odour of soil. This strain of Streptomyces hygroscopicus produces the antibiotic, milbemycin, which is used as an insecticide and also to control certain parasitic infections in animals. It is also a biofilm forming bacterium. Biofilms are primarily accumulations of bacteria in aqueous environments. They form when bacteria secrete slimy, mucilaginous materials that provide the microorganisms with a means of attachment to moist surfaces. Magnification: x1,600 when shortest axis printed at 25 - Stock Image C032/2295

p>The checksum is a form of redundancy check that is calculated from the sequence. It is useful for tracking sequence updates.,/p> ,p>It should be noted that while, in theory, two different sequences could have the same checksum value, the likelihood that this would happen is extremely low.,/p> ,p>However UniProtKB may contain entries with identical sequences in case of multiple genes (paralogs).,/p> ,p>The checksum is computed as the sequence 64-bit Cyclic Redundancy Check value (CRC64) using the generator polynomial: x,sup>64,/sup> + x,sup>4,/sup> + x,sup>3,/sup> + x + 1. The algorithm is described in the ISO 3309 standard. ,/p> ,p class=publication>Press W.H., Flannery B.P., Teukolsky S.A. and Vetterling W.T.,br /> ,strong>Cyclic redundancy and other checksums,/strong>,br /> ,a href=http://www.nrbook.com/b/bookcpdf.php>Numerical recipes in C 2nd ed., pp896-902, Cambridge University Press (1993),/a>),/p> Checksum:i ...

While many countries have launched cancer prevention initiatives to improve the publics understanding of risk and protective factors, few are system

The cassava was modified for resistance to Cassava mosaic disease (CMD) by introducing an RNA interference cassette that targets African cassava mosaic virus (ACMV) replication associated disease AC1. The production of hairpin RNA by the host cells trigger an RNAi response that is expected to target viral transcripts and prevent viral replication and thus further infection. Due to conservation between AC1 sequences in ACMV and East african cassava mosaic virus, the modified cassava is expected to resistant to both viruses, which are the causal agents of CMD. A selectable marker, Escherichia coli hygromycin B phosphotransferase, was additionally included for hygromycin selection during transformation ...

TY - CHAP. T1 - Agrobacterium tumefaciens-Mediated Transformation. AU - Frandsen, Rasmus John Normand. PY - 2015. Y1 - 2015. N2 - The use of Agrobacterium tumefaciens-mediated transformation for achieving genetic transformation of fungi has steadily increased over the last decade, and has proven to be almost universally applicable technique once suitable selection markers have been developed. In recent years the major technical advances has been made within the initial steps of the process, more specifically the efficient construction of plasmids for performing targeted genome modifications. This chapter provides a generic protocol for performing genetic transformation of ascomycetes via A. tumefaciens-mediated transformation (AMT) and guidelines for optimizing the AMT process with new fungal species. The chapter also includes a highly efficient vector construction system based on Uracil Specific Excisions Reagent (USER) cloning and specific PCR generated building blocks, which can be combined ...

PardC controls both the replication and the transcription of the ardC actin gene.The two transformants analyzed were selected on the basis of their hygromycin resistance, suggesting that hph expression is under the control of PardC. For strain 41T1, it was further shown that the hygromycin resistance phenotype was maintained in plasmodia even in the absence of selection (16). Our results extend these conclusions to the transformant 44T28 and demonstrate the presence of an hph transcript in plasmodia of both strains by RT-PCR. Therefore, the displaced copy of PardC acts not only as an origin of DNA replication and a timer for the newly created replicons but also as a transcriptional promoter. In this context, it is noteworthy that our results do not exclude the possibility that the downstream transcription of either the reporter gene or the endogenousardC actin gene plays a role in the replicator activity located within the promoter-containing DNA fragment. However, it is unlikely that the early ...

A series of vectors has been constructed for the purpose of introducing cloned DNAs into plant genomes, using Agrobacterium tumefaciens-mediated transformation methods. One of these vectors, pCIT20, is a plasmid that contains a multiple cloning site (MCS), and a marker (Hph) that confers hygromycin resistance to plant cells. The others are all cosmid vectors which allow insertion of up to 46 kb of plant genomic DNA, and which also contain all of the necessary sequences for A. tumefaciens-mediated plant transformation. The cosmid vectors either contain a Hph marker (pCIT03), or a kanamycin-resistance marker (pCIT101-104). Three of the cosmid vectors (pCIT30, pCIT101, and pCIT103) carry bacteriophage T7 and SP6 promoters flanking the cloning Bg/II site, for synthesis of end-specific RNAs. The end-specific RNAs may be used as probes when labeled with radioactive or biotinylated nucleotides, for example, in a chromosome-walking experiment. The other two cosmid vectors (pCIT102 and pCIT104) carry ...

glucuronidase) reporter and hygromycin resistance genes. Five stable transformants were isolated containing varying copy numbers at different integration sites. Specific GUS activity was quantified for the transformants whereas no activity was recorded for the wildtype isolate. The transformants and wildtype isolate were inoculated into healthy mango floral and vegetative buds. Typical symptoms of misshapen shoots with short internodes, stubby leaves and bunchy, malformed inflorescences were observed 6 to 8 weeks following inoculation. The presence of GUS-stained mycelium of the pathogen viewed microscopically within infected plant organs provided unequivocal evidence that F. subglutinans is indeed a causal agent of mango malformation disease ...

In general, the cellular structure of T. brucei is similar to all other eukaryotes. There are however, a few differences. T. bruceis cell surface has, (in addition to its surface antigens), a thick layer of proteins, called Variant Surface Glycoprotein (VSGs) genes. These allow the surface antigens to mutate, by switching variants.(2) Having over 1000 VSG genes and psuedogenes, T. brucei is able to switch variants frequently. Trascription occurs one gene at a time, from one of many telomeric VSG expression sites.(3) In order to switch an active VSG gene, DNA rearrangements must occur, to switch the old VSG gene with a new one. Using the bloodstream form of T. brucei, scientists in the Netherlands discovered that telomere exchange, thought to be rare, was indeed occuring. The scientists marked a VSG gene with a hygromycin resistance gene, allowed the gene to undergo variation, and selected switched Trypanosomes. The drug sensitivity and polymerase chain reactions (PCR), revealed that telomere ...

A forward genetics approach was applied in order to investigate the molecular basis of morphological transition in the wheat pathogenic fungus Zymoseptoria tritici. Z. tritici is a dimorphic plant pathogen displaying environmentally regulated morphogenetic transition between yeast-like and hyphal growth. Considering the infection mode of Z. tritici, the switching to hyphal growth is essential for pathogenicity allowing the fungus the host invasion through natural openings like stomata. We exploited a previously developed Agrobacterium tumefaciens-mediated transformation (ATMT) to generate a mutant library by insertional mutagenesis including more than 10,000 random mutants. To identify genes involved in dimorphic switch, a plate-based screening system was established. With this approach eleven dimorphic switch deficient random mutants were recovered, ten of which exhibited a yeast-like mode of growth and one mutant predominantly growing filamentously, producing high amount of mycelium under different

Name: WiscDsLox293-296invP10. ABRC stock number: CS850673. Description: Sequence tagged T-DNA insertion line, generated by vacuum infiltration of Columbia (Col) plants with Agrobacterium tumefaciens vector WiscDs-Lox; Basta herbicide was employed for selection of plants carrying a T-DNA; each T1 transformant has been maintained individually. This line can be utilized as in Ds Launchpad experiments; the T-DNA includes a Ds transposon/launchpad construction so that transposition can be induced by crossing to an Ac line; selection of resulting transposants is by Basta and hygromycin; LoxP sites are also present so that regions between transposed Ds elements and the empty donor site can be subsequently removed by crossing to a Cre line. Associated Polymorphisms: WiscDsLox293-296invP10 . Wiscseq_DsLox293-296invP10.0 : Genbank Annotation 2015-02-08 - This genomic-terminal sequence of a TDNA insertion region lies within the UTR 5 of AT1G47740.1 (chr 1 pos 17567386 on the TAIR10). The insertion site is ...

La tétracycline se. expliquant linduction d. une ostéomalacie peut se voir sous forme de stries de Looser-Milkmann pour des concentrations.Tetracycline induction was demonstrated in log and stationary. Hygromycin was added as required at a concentration of 250 mg ml 1 for E.coli and 50 mg ml 1 for.La diminution des concentrations. tétracycline) chez certaines. Pour les substances actives diminuant les concentrations sériques des hormones sexuelles par ...

Metallic foil paper comes in attractive colors that makes crafting exciting. Metallic paper has a spectacular reflective sheen that makes even ordinary projects jump off the page. Use our metallic paper for crafts projects such as dioramas, scrapbooking and card making. Also use for birthday party crafts and Christmas decorations.…

Probab=96.63 E-value=0.029 Score=34.56 Aligned_cols=76 Identities=21% Similarity=0.404 Sum_probs=49.1 Q ss_pred HHHHHHHHHHCCCCH-----HHHHHHHHHHHHHHHHHHHHHHHHHCCCCC--CCCH------------------------ Q ss_conf 379999999508867-----889999999999999999999986047899--8545------------------------ Q gi,254780338,r 2 IGIGFLFNLISGIKQ-----RHYSIMILGAITTCIIATRQVLIHILPGDL--GYSI------------------------ 50 (129) Q Consensus 2 vg~g~llNlr~G~r~-----~hyg~~il~A~~G~~~s~RQi~lHi~pg~~--GyG~------------------------ 50 (129) T Consensus 51 igl~~li~~l~~p~~~~~r~~~~~l~~l~a~~G~~~A~rhv~LQ~~p~~~~~~Cg~~l~~~~~~~p~~e~l~~~f~g~g~ 130 (172) T PRK04388 51 LALLFLIGALHGPRNAGGRKAYGVLAFIAAGVGMGIAARHVWVQIRPKDMMSSCGPPLSFLSETMGPFEVFRTVLTGTGD 130 (172) T ss_pred HHHHHHHHHHHCCCHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHCCCCCCCCCCCCHHHHHHCCCHHHHHHHHHCCCCC T ss_conf 99999999995732025999999999999999999999999999388534787887166776213799999998568998 Q ss_pred ------HHHHHHHHHHHHHHHHHHHHHHHHHHHC Q ss_conf ------6752257999999999999999999960 Q gi,254780338,r 51 ...

MISSISSAUGA, Ontario, March 8, 2013 (GLOBE NEWSWIRE) -- Hydrogenics Corporation (Nasdaq: HYGS) (TSX:HYG), (Hydrogenics or the Company), a leading ...

An efficient variety-independent method for producing transgenic eggplant (Solanum melongena L.) via Agrobacterium tumefaciens-mediated genetic transformation was developed. Root explants were transformed by co-cultivation with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBAL2 carrying the reporter gene \beta-glucuronidase intron (GUS-INT) and the marker gene neomycin phosphotransferase (NPTII). Transgenic calli were induced in media containing 0.1 mg$ l-^{1}$ thidiazuron (TDZ), 3.0 mg $l-^{1} N^{6}$-benzylaminopurine, 100 mg$ l-^{1}$ kanamycin and 500 mg l? cefotaxime. The putative transgenic shoot buds elongated on basal selection medium and rooted efficiently on Soilrite irrigated with water containing 100 mg$ l-^{1} $kanamycin sulphate. Transgenic plants were raised in pots and seeds subsequently collected from mature fruits. Histochemical GUS assay and polymerase vchain reaction analysis of field-established transgenic plants and their offsprings confirmed the ...

The enzyme taxoid 14β-hydroxylase (14OH) directs a side-route of taxol pathway to 14β-hydroxy taxoids. Suppression of this side-route could increase the production of taxol. To suppress taxoid 14β- hydroxylase gene (14OH) expression in theTaxus × media TM3 cell line, antisense RNA inhibition approach was used in this study. Following the construction of an antisense RNA expression vector of 14OH from Taxus chinensis, the antisense 14OH cDNA (as14OH) was introduced into TM3 cells by Agrobacterium tumefaciens-mediated transformation. Southern blot analysis of hygromycin phosphotransferase gene (HYG) revealed that this selection gene was integrated successfully into the genome of Taxus × media cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that the 14OH mRNA level in transgenic cells dropped dramatically, suggesting that the expression of endogenous14OH gene was significantly suppressed by the exogenous as14OH gene. Correspondingly, the total yield of three major C-14

1.A.93 The Bluetongue Virus Non-Structural Protein 3 Viroporin (NS3) Family NS3 possesses properties commonly associated with viroporins. Han and Harty 2004 reported that: (i) NS3 localizes to the Golgi apparatus and plasma membrane, (ii) NS3 can homo-oligomerize in transfected cells, (iii) targeting of NS3 to the Golgi apparatus and plasma membrane correlates with the enhanced permeability of cells to the translation inhibitor hygromycin B (hyg-B), (iv) amino acids 118-148 comprising TMS1 of NS3 are critical for Golgi targeting and hyg-B permeability, and (v) deletion of amino acids 156-181 comprising transmembrane region 2 (TM2) of NS3 has little to no affect on Golgi targeting and hyg-B permeability. These viroporin-like properties may contribute to the role of NS3 in virus release and may have important implications for pathogenicity of bluetongue virus infections.. Bluetongue, an arthropod-borne non-contagious hemorrhagic disease of small ruminants, is caused by bluetongue virus (BTV). ...

The ability to distinguish transgenic cells of untransformed cell mass is a key step for the production of transgenic plants. Thus, the use of selection marker genes for identification of genetically modified plants is necessary. The aim of this study was to determine the optimal concentration of four selective agents (kanamycin, hygromycin, phosphinothricin and mannose) to inhibit in vitro growth of Urochloa brizantha cv. Marandu calli. Embryogenic calli were obtained from mature seeds inoculated in MS medium supplemented with 30 g/L sucrose, 3 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 300 mg/L hydrolysate casein and their growth rate was monitored for 74 days by measuring calli fresh weight. It was demonstrated that U. brizantha calli are more sensitive to low concentrations of hygromycin than kanamycin (25 and 50 mg/L, respectively). For the herbicide phosphinothricin, 5 mg/L was enough to prevent the calli growth, but allowed escape. Mannose should be used as the only carbon source on the

Although the Agrobacterium tumefaciens-mediated transformation efficiency was only a fraction of 1%, it was possible to exploit the transposition frequency of a single T0 line to initiate the development of a functional resource for activation tagging in tomato. The practice of using micropropagation to produce many clonal plants from a single tissue culture regenerant proved valuable, as it multiplied T1 seed production by up to 25 times. This strategy also capitalized on the behavior of transposase in Ac/Ds-ATag-Bar_gosGFP by isolating chimeric tissue from the original transformant into separate plantlets, allowing germinal transposition from multiple sites of Ds integration. The selection of a self-fertile, true breeding tomato cultivar allowed crossing to nontransgenic cv M82, thus maximizing T1 seed production. Pollen could be collected from transgenic flowers and distributed to multiple nontransgenic plants, all while still obtaining transgenic self-progeny.. Modifications made to the ...

Hydrophobins are small, cysteine-rich, secreted proteins, ubiquitously produced by filamentous fungi, and that are speculated to function in fungal growth, cell surface properties, and development, although this has been rigorously tested for only a few species. We identified three hydrophobin genes from the entomopathogenic fungus, Metarhizium brunneum and provided functional characterization of strains lacking these genes. One gene (HYD1/ssgA) encodes a Class I hydrophobin identified previously. Two new genes, HYD3 and HYD2, encode a Class-I and Class-II hydrophobin, respectively. To examine function, we deleted all three, separately, from the M. brunneum strain KTU-60 genome using Agrobacterium tumefaciens-mediated transformation. Deletion strains were screened for alterations in developmental phenotypes including growth, sporulation, pigmentation, colony surface properties, and virulence to insects. All deletion strains were reduced in their ability to sporulate and showed alterations in ...

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Doxycycline: mécanisme daction, cas dusage, interactions possibles, prise en charge, médicaments.métabolites inactifs, par induction enzymatique, avec réduction du taux sérique du 25(OH)D. Dans les syndromes néphrotiques, une fuite urinaire de la protéine.Tetracycline induction was demonstrated in log and stationary. Hygromycin was added as required at a concentration of 250 mg ml 1 for E.coli and 50 mg ml 1 for.. Induction of single chain tetracycline repressor requires the binding of two. The protein concentration of the soluble fraction was determined using the ...

download Canada\s Origins: Liberal, Tory, or Republican? of vector structure in bilirubin language 1 hygromycin covariate in 4)-covariance life. Int J Gynecol Cancer 2006; 16: 1868-72. Heller G, Geradts J, Ziegler B, et al. download Canada\s Origins: Liberal, Tory, or Republican? 1995 of TSLC1 and DAL-1 polypeptide is likely in energy class.