A heat shock following electroporation induces highly efficient transformation of Corynebacterium glutamicum with xenogeneic plasmid DNA

被引：448

作者：

van der Rest, ME ^{[1
]}

Lange, C ^{[1
]}

Molenaar, D ^{[1
]}

机构：

[1] Univ Dusseldorf, Biotechnol Zent Lab, D-40225 Dusseldorf, Germany

来源：

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | 1999年 / 52卷 / 04期

关键词：

D O I：

10.1007/s002530051557

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An improved method for the electrotransformation of wild-type Corynebacterium glutamicum? (ATCC 13032) is described. The two crucial alterations to previously developed methods are: cultivation of cells used for electrotransformation at 18 degrees C instead of 30 degrees C, and application of a heat shock immediately following electrotransformation. Cells cultivated at sub optimal temperature have a 100-fold improved transformation efficiency (10(8) cfu mu g(-1)) for syngeneic DNA (DNA isolated from the same species). A heat shock applied to these cells following electroporation improved the transformation efficiency for xenogeneic DNA (DNA isolated from a different species). In combination, low cultivation temperature and heat shock act synergistically and increased the transformation efficiency by four orders of magnitude to 2.5 x 10(6) cfu mu g(-1) xenogeneic DNA. The method was used to generate gene disruptions in C. glutamicum.

引用

页码：541 / 545

页数：5

共 18 条

[11]

Sambrook J., 2002, MOL CLONING LAB MANU

[12]

SCHAFER A, 1994, J BACTERIOL, V176, P7309

[13]

SCHAFER A, 1994, APPL ENVIRON MICROB, V60, P756

[14] SMALL MOBILIZABLE MULTIPURPOSE CLONING VECTORS DERIVED FROM THE ESCHERICHIA-COLI PLASMIDS PK18 AND PK19 - SELECTION OF DEFINED DELETIONS IN THE CHROMOSOME OF CORYNEBACTERIUM-GLUTAMICUM [J].

SCHAFER, A ;

TAUCH, A ;

JAGER, W ;

KALINOWSKI, J ;

THIERBACH, G ;

PUHLER, A .

GENE, 1994, 145 (01) :69-73

[15] MANIPULATION OF CORYNEBACTERIUM-GLUTAMICUM BY GENE DISRUPTION AND REPLACEMENT [J].

SCHWARZER, A ;

PUHLER, A .

BIO-TECHNOLOGY, 1991, 9 (01) :84-87

[16] INCREASE IN PLASMID TRANSFORMATION EFFICIENCY IN SOS-INDUCED ESCHERICHIA-COLI-CELLS [J].

VERICAT, JA ;

GUERRERO, R ;

BARBE, J .

MOLECULAR & GENERAL GENETICS, 1988, 211 (03) :526-530

[17] REPLACEMENT RECOMBINATION IN CORYNEFORM BACTERIA - HIGH-EFFICIENCY INTEGRATION REQUIREMENT FOR NONMETHYLATED PLASMID DNA [J].

VERTES, AA ;

HATAKEYAMA, K ;

INUI, M ;

KOBAYASHI, M ;

KURUSU, Y ;

YUKAWA, H .

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1993, 57 (12) :2036-2038

[18] A new type of transporter with a new type of cellular function: L-lysine export from Corynebacterium glutamicum [J].

Vrljic, M ;

Sahm, H ;

Eggeling, L .

MOLECULAR MICROBIOLOGY, 1996, 22 (05) :815-826

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