ELECTROTRANSFORMATION OF THIOBACILLUS-FERROOXIDANS WITH PLASMIDS CONTAINING A MER DETERMINANT

被引：50

作者：

KUSANO, T ^{[1
]}

SUGAWARA, K ^{[1
]}

INOUE, C ^{[1
]}

TAKESHIMA, T ^{[1
]}

NUMATA, M ^{[1
]}

SHIRATORI, T ^{[1
]}

机构：

[1] DOWA MIN CO LTD,CENT RES LAB,HACHIOJI,TOKYO 193,JAPAN

来源：

JOURNAL OF BACTERIOLOGY | 1992年 / 174卷 / 20期

关键词：

D O I：

10.1128/JB.174.20.6617-6623.1992

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The mer operon from a strain of Thiobacillus ferrooxidans (C. Inoue, K. Sugawara, and T. Kusano, Mol. Microbiol. 5:2707-2718, 1991) consists of the regulatory gene merR and an operator-promoter region followed by merC and merA structural genes and differs from other known gram-negative mer operons. We have constructed four potential shuttle plasmids composed of a T. ferrooxidans-borne cryptic plasmid, a pUC18 plasmid, and the above-mentioned mer determinant as a selectable marker. Mercury ion-sensitive T. ferrooxidans strains were electroporated with constructed plasmids, and one strain, Y4-3 (of 30 independent strains tested), was found to have a transformation efficiency of 120 to 200 mercury-resistant colonies per mug of plasmid DNA. This recipient strain was confirmed to be T. ferrooxidans by physiological, morphological, and chemotaxonomical data. The transformants carried a plasmid with no physical rearrangements through 25 passages under no selective pressure. Cell extracts showed mercury ion-dependent NADPH oxidation activity.

引用

页码：6617 / 6623

页数：7

共 45 条

[1] ACTIVITY OF THE HYBRID TRP-LAC(TAC) PROMOTER OF ESCHERICHIA-COLI IN PSEUDOMONAS-PUTIDA - CONSTRUCTION OF BROAD-HOST-RANGE, CONTROLLED-EXPRESSION VECTORS [J].