Directional Cloning into Plasmid Vectors

Sambrook, Joseph; Russell, David W.

doi:10.1101/pdb.prot3919

Cited by 323 publications

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“…Total genomic DNA was purified and used to construct genomic libraries using a variety of single restriction enzymes (Sambrook et al, 1989) and the cloning vector pSU18 (Martinez et al, 1988). The genomic libraries were used to transform E. coli DH5a (Invitrogen) to chloramphenicol resistance (30 mg ml 21 ) and recombinants containing a cloned Mini-Tn903-Km R cassette were selected with kanamycin (25 mg ml…”

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confidence: 99%

Induction of -lactamase production in Aeromonas hydrophila is responsive to -lactam-mediated changes in peptidoglycan composition

et al. 2010

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We have studied the mechanism by which b-lactam challenge leads to b-lactamase induction in Aeromonas hydrophila through transposon-insertion mutagenesis. Disruption of the DDcarboxypeptidases/endopeptidases, penicillin-binding protein 4 or BlrY leads to elevated monomer-disaccharide-pentapeptide levels in A. hydrophila peptidoglycan and concomitant overproduction of b-lactamase through activation of the BlrAB two-component regulatory system. During b-lactam challenge, monomer-disaccharide-pentapeptide levels increase proportionately with b-lactamase production and b-lactamase induction is inhibited by vancomycin, which binds muro-pentapeptides. Taken together, these data strongly suggest that the Aeromonas spp. blactamase regulatory sensor kinase, BlrB, responds to the concentration of monomerdisaccharide-pentapeptide in peptidoglycan.

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confidence: 99%

Induction of -lactamase production in Aeromonas hydrophila is responsive to -lactam-mediated changes in peptidoglycan composition

et al. 2010

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“…The markers were also checked in 12% polyacrylamide gel electrophoresis (Atto, Japan, Vertical unit). The polyacrylamide gels were visualized by silver staining (20 ml methanol, 1 ml glacial acetic acid, 0.2 g AgNO 3 179 ml distilled water) after fixing in a solution of 20 ml methanol, 1 ml glacial acetic acid, 179 ml distilled water, followed by destaining in a solution of 600 µl formaldehyde, 199.4 ml distilled water, 5.1 g NaOH, and photographed in Bio-Rad gel documentation system (Sambrook and Russell 2001).…”

Section: Molecular Analysismentioning

confidence: 99%

Transferability and Polymorphism Between Group 7 Chromosome Specific Simple Sequence Repeat (SSR) Markers of Bread Wheat and Its Related Non-ProgenitorAegilopsSpecies

Kumar

Verma

Kundu

et al. 2016

Journal of Crop Improvement

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Simple sequence repeat (SSR) markers are highly efficient for genetic mapping and molecular breeding in crop plants. Chromosomes 7S and 7U of related non-progenitor Aegilops species possess superior genetic systems for high grain iron and zinc content. This study was undertaken to conduct a comprehensive analysis of transferability and polymorphism among group-7 anchored SSR markers of bread wheat (Triticum aestivum L.) to Aegilops species for wheat improvement. The study revealed 77% transferability of group 7-specific SSR markers of bread wheat to 7U/7S chromosomes of Aegilops species. More than 80% of the 7D specific markers were found to be transferable with a high level of polymorphism. The transferability to 7S and 7U genome suggested higher similarity between bread wheat genome(s) and S genome as compared with U genome of Aegilops species. These polymorphic markers are highly informative, robust and cost-effective for molecular breeding and could be further utilized for identification of introgression/transfer(s) of 7S/7U chromosomal fragments in bread wheat genome through wide hybridization. ARTICLE HISTORY

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“…In a replica gel molecular weight marker proteins were run. After electrophoresis, the sample gel was dried and phosphorimaged by Typhoon 9210 (GE Healthcare) and the gel with marker proteins was stained with coommassie brilliant blue, as described in [27].…”

Section: -D Gel Electrophoresismentioning

confidence: 99%

Calcium Chloride Made E. coli Competent for Uptake of Extraneous DNA Through Overproduction of OmpC Protein

et al. 2012

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In the standard method of transformation of Escherichia coli with extraneous DNA, cells are made competent for DNA uptake by incubating in ice-cold 100 mM CaCl(2). Analysis of the whole protein profile of CaCl(2)-treated E. coli cells by the techniques of one- and two-dimensional gel electrophoresis, MALDI-MS and immunoprecipitation revealed overproduction of outer membrane proteins OmpC, OmpA and heat-shock protein GroEL. In parity, transformation efficiency of E. coli ompC mutant by plasmid pUC19 DNA was found to be about 40 % lower than that of the wild type strain. Moreover, in E. coli cells containing groEL-bearing plasmid, induction of GroEL caused simultaneous overproduction of OmpC. On the other hand, less OmpC was synthesized in E. coli groEL mutant compared to its wild type counterpart, by CaCl(2)-shock. From these results it can be suggested that in the process of CaCl(2)-mediated generation of competence, the heat-shock chaperone GroEL has specific role in DNA entry into the cell, possibly through the overproduced OmpC and OmpA porins.

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Directional Cloning into Plasmid Vectors

Cited by 323 publications

References 0 publications

Induction of -lactamase production in Aeromonas hydrophila is responsive to -lactam-mediated changes in peptidoglycan composition

Induction of -lactamase production in Aeromonas hydrophila is responsive to -lactam-mediated changes in peptidoglycan composition

Transferability and Polymorphism Between Group 7 Chromosome Specific Simple Sequence Repeat (SSR) Markers of Bread Wheat and Its Related Non-ProgenitorAegilopsSpecies

Calcium Chloride Made E. coli Competent for Uptake of Extraneous DNA Through Overproduction of OmpC Protein

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