A novel episomal shuttle vector for transformation of Cryptococcus neoformans with the ccdB gene as a positive selection marker in bacteria

被引：29

作者：

Mondon, P ^{[1
]}

Chang, YC ^{[1
]}

Varma, A ^{[1
]}

Kwon-Chung, KJ ^{[1
]}

机构：

[1] NIAID, LCI, Mol Microbiol Sect, NIH, Bethesda, MD 20892 USA

来源：

FEMS MICROBIOLOGY LETTERS | 2000年 / 187卷 / 01期

关键词：

positive selection; vector; transformation; Cryptococcus neoformans;

D O I：

10.1111/j.1574-6968.2000.tb09134.x

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

We report the engineering of a new shuttle vector featuring its episomal maintenance in Cryptococcus neoformans and the lethal Escherichia coli ccdB gene for positive selection in bacteria. Telomere-like sequences from C. neoformans and the STAB fragment confer episomal maintenance to the vector (pPM8) upon transformation in C. neoformans. The vector generated high transformation frequencies and each transformant was estimated to harbor thirty copies of the plasmid. The plasmids recovered in E. coli from the C. neoformans transformants showed no evidence of rearrangement. This construct will be very useful for cloning and studying the regulation of genes in C. neoformans. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

引用

页码：41 / 45

页数：5

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