An improved protocol for electrotransformation of Corynebacterium pseudotuberculosis

被引：24

作者：

Dorella, F. A.

Estevam, E. M.

Cardoso, P. G.

Savassi, B. M.

Oliveira, S. C.

Azevedo, V.

Miyoshi, A. ^{[1
]}

机构：

[1] Univ Fed Minas Gerais, Lab Genet Celular & Mol, Inst Ciencias Biol, Belo Horizonte, MG, Brazil

[2] Univ Fed Minas Gerais, Lab Imunol Doencas Infecciosas, Inst Ciencias Biol, Belo Horizonte, MG, Brazil

来源：

VETERINARY MICROBIOLOGY | 2006年 / 114卷 / 3-4期

关键词：

Corynebacterium pseudotuberculosis; plasmid transformation; electroporation;

D O I：

10.1016/j.vetmic.2005.12.010

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

We developed an improved protocol for the electrotransformation of Corynebacterium pseudotuberculosis, testing variations of parameters in the procedures that are routinely used for the preparation of electrocompetent cells of this species, including (i) culture conditions, (ii) cell growth phase, (iii) electroporation solutions and (iv) quantity of plasmid DNA. We obtained the greatest efficiency of transformation when the cells were grown until the stationary phase and then washed with 10% glycerol electroporation solution. The transformation efficiency was inversely proportional to the quantity of plasmid DNA. The transformation efficiency reached 10(5) colony-forming units (cfu)/mu g plasmid DNA. This protocol would be useful for genetic studies of C. pseudotuberculosis. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：298 / 303

页数：6

共 26 条

[1] Growth of Escherichia coli on medium containing glycine increases transformation efficiency [J].