ELECTROTRANSFORMATION IN SALMONELLA-TYPHIMURIUM LT2

被引:51
|
作者
BINOTTO, J [1 ]
MACLACHLAN, PR [1 ]
SANDERSON, KE [1 ]
机构
[1] UNIV CALGARY,DEPT BIOL SCI,CALGARY T2N 1N4,ALBERTA,CANADA
关键词
ELECTROPORATION; SALMONELLA; GENETICS; TRANSFORMATION;
D O I
10.1139/m91-078
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Electroporation gives high efficiency of transformation in Salmonella typhimurium LT2, yielding 10(8)-10(9) electrotransformants per microgram of pBR322 DNA. Lipopolysaccharide (LPS) composition has little influence on electrotransformation efficiency by electroporation, unlike Ca2+ shock methods, which give ca. 10(6) transformants/mu-g DNA with strains with Rc or Rd2 LPS, 10(4) transformants with most smooth and rough strains, and 10(2) transformants with strains with Re LPS. Thus cell envelope properties are less crucial in electrotransformation than in Ca2+ shock methods. The reciprocal restriction barrier between Escherichia coli K-12 and S. typhimurium LT2 reduces electrotransformation by ca. 100-fold, but host-restriction mutants reduce or eliminate the barrier.
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页码:474 / 477
页数:4
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