Suzuki S, Fukagawa T
Department of Aquaculture, Kochi University, Japan.
J Ind Microbiol. 1995 Feb;14(2):154-8. doi: 10.1007/BF01569897.
A tributyltin chloride (TBTCl)-resistant bacterium, Alteromonas sp. M-1, was isolated from coastal seawater. This bacterium grew in medium containing 125 microM TBTCl. TBTCl added to the medium was taken up by this bacterium, however, the amount of TBTCl in the cellular fraction was low after the logarithmic phase, suggesting the existence of a TBTCl-efflux system. A genetic library was constructed using plasmid vector pUC 19. Three positive clones were obtained, by which E. coli was transformed to TBTCl resistance. Of the three clones, the shortest fragment from HindIII-library was analyzed. This fragment was 1.8 kb long and contained one complete open reading frame. The predicted amino acid sequence of this open reading frame had a homologous domain to transglycosylases of bacteriophage and E. coli. TBTCl-tolerant marine bacteria other than Alteromonas sp. M-1 were obtained from natural seawater to which TBTCl was added. DNA-DNA hybridization was performed between the three cloned fragments from Alteromonas sp. M-1 and chromosomal DNA of the TBTCl-tolerant bacteria. Some strains hybridized with the fragments and some did not, suggesting that several genes are responsible for TBTCl tolerance.
从沿海海水中分离出一株对三丁基氯化锡(TBTCl)具有抗性的细菌——交替单胞菌属M-1菌株。该细菌能在含有125微摩尔/升TBTCl的培养基中生长。添加到培养基中的TBTCl会被该细菌摄取,然而,在对数生长期后,细胞组分中TBTCl的含量较低,这表明存在TBTCl外排系统。使用质粒载体pUC 19构建了一个基因文库。获得了三个阳性克隆,通过它们将大肠杆菌转化为对TBTCl具有抗性。对这三个克隆中来自HindIII文库的最短片段进行了分析。该片段长1.8 kb,包含一个完整的开放阅读框。这个开放阅读框的预测氨基酸序列与噬菌体和大肠杆菌的转糖基酶具有同源结构域。除交替单胞菌属M-1菌株外,还从添加了TBTCl的天然海水中获得了对TBTCl具有耐受性的海洋细菌。对来自交替单胞菌属M-1菌株的三个克隆片段与对TBTCl具有耐受性的细菌的染色体DNA进行了DNA-DNA杂交。一些菌株与这些片段杂交,而一些则没有,这表明有几个基因与TBTCl耐受性有关。