Open AccessHigh efficiency electroporation of intact Corynebacterium glutamicum cells
Author(s) -
Liebl W.,
Bayerl A.,
Schein B.,
Stillner U.,
Schleifer K.H.
Publication year - 1989
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1989.tb03677.x
Subject(s) - corynebacterium glutamicum , electroporation , plasmid , transformation (genetics) , dna ligase , transformation efficiency , dna , biology , corynebacterium , bacteria , plasmid preparation , exogenous dna , microbiology and biotechnology , chemistry , biochemistry , gene , genetics , pbr322 , agrobacterium
High‐frequency electroporation of whole Corynebacterium glutamicum cells without enzymatic pretreatment was achieved. Under optimized conditions concerning growth stage, washing of cells, cell concentration and pulse parameters transformation efficiencies of far more than 10 7 transformants per μg pWST4B plasmid DNA were reached. Using electroporation, linearised and subsequently religated plasmid as well as chimeric ligase reaction products were directly introduced into C. glutamicum with reasonable efficiencies. Electrotransformation efficiency was reduced about 10 5 ‐fold for plasmid DNA cycled through E. coli JM83. Restriction deficient mutants of C. glutamicum were isolated which could be efficiently transformed with foreign DNA.