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Transformation of Vibrio cholerae by plasmid DNA

IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata

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Field Value
 
Title Transformation of Vibrio cholerae by plasmid DNA
 
Creator Panda, Dibyendu K
Dasgupta, Ujjata
Das, Jyotirmoy
 
Subject Infectious Diseases and Immunology
 
Description The lack of an eficient transformation system in Vibrio c~oler~e was a handicap in the genetic manipulation of this important human pathogen. Since V. cholerae cells secrete DNases, this may interfere with the uptake of DNA. The present report describes the approaches taken for transforming V’. cholerae cells with plasmid DNA, by overcoming this DNase barrier. The partial success of transforming DNase-negative mutants confirmed the role of DNase in the nontransformability of the wild-type cells. Successful transformation was carried out following removal of DNases from the periplasmic space. This was achieved by treating the cells with Mg2 + and Ca* + ions to allow the DNase to be released, and then holding them under conditions where the remaining DNase activity was minimized before adding DNA to the competent cells. Transformation effxciencies of the order of 10 5 per recipient cell were observed.
 
Date 1991
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/2291/1/GENE__V._105__(_1_)_107%2D111_;1991.pdf
Panda, Dibyendu K and Dasgupta, Ujjata and Das, Jyotirmoy (1991) Transformation of Vibrio cholerae by plasmid DNA. Gene, 105 (1). pp. 107-111.
 
Relation http://www.eprints.iicb.res.in/2291/