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High-frequency electroporation and maintenance of pUC- and pBR-based cloning vectors in Pseudomonas stutzeri.

作者信息

Pemberton J M, Penfold R J

机构信息

Microbiology Department, University of Queensland, St. Lucia, Brisbane, Australia.

出版信息

Curr Microbiol. 1992 Jul;25(1):25-9. doi: 10.1007/BF01570078.

DOI:10.1007/BF01570078
PMID:1369188
Abstract

A number of Escherichia coli cloning vectors, based on ColE1-like replicons, were shown to be maintained in Pseudomonas stutzeri ATCC 17588. A restrictionless mutant of P. stutzeri was isolated, and this strain was used to develop an efficient electroporation system. With the E. coli cloning vector pHSG298, transformation frequencies of up to 2 x 10(7) transformants/micrograms DNA were achieved. This frequency is comparable to that obtained for CaCl2-mediated transformation of E. coli; thus, direct cloning of DNA into P. stutzeri is feasible. As will be discussed, this may prove useful for cloning DNA from high mol% G + C genera in cases in which E. coli is not a suitable heterologous cloning host.

摘要

相似文献

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引用本文的文献

1
Pseudomonas stutzeri has two closely related pilA genes (Type IV pilus structural protein) with opposite influences on natural genetic transformation.施氏假单胞菌有两个密切相关的pilA基因(IV型菌毛结构蛋白),它们对自然遗传转化有着相反的影响。
J Bacteriol. 2001 Apr;183(7):2359-66. doi: 10.1128/JB.183.7.2359-2366.2001.
2
Type IV pilus genes pilA and pilC of Pseudomonas stutzeri are required for natural genetic transformation, and pilA can be replaced by corresponding genes from nontransformable species.斯氏假单胞菌的IV型菌毛基因pilA和pilC是自然遗传转化所必需的,并且pilA可以被来自非转化物种的相应基因所取代。
J Bacteriol. 2000 Apr;182(8):2184-90. doi: 10.1128/JB.182.8.2184-2190.2000.
3

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High-copy-number and low-copy-number plasmid vectors for lacZ alpha-complementation and chloramphenicol- or kanamycin-resistance selection.用于lacZα互补以及氯霉素或卡那霉素抗性筛选的高拷贝数和低拷贝数质粒载体。
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