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KMID : 0545120070170010162
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 1 p.162 ~ p.167
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Construction of Conjugative Gene Transfer System Between E. coli and Moderately Thermophilic, Extremely Acidophilic Acidithiobacillus caldus MTH-04
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Liu Xiangmei
Lin Jianqun
Zhang Zheng
Bian Jiang
Zhao Qing
Liu Ying
Lin Jianqiang
Yan Wangming
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Abstract
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A genetic transfer system for introducing foreign genes to biomining microorganisms is urgently needed. Thus, a conjugative gene transfer system was investigated for a moderately thermophilic, extremely acidophilic biomining bacterium, Acidithiobacillus caldus MTH-04. The broad-hostrange IncP plasmids RP4 and R68.45 were transferred directly into A. caldus MTH-04 from Escherichia coli by conjugation at relatively high frequencies. Additionally the broad-hostrange IncQ plasmids pJRD215, pVLT33, and pVLT35 were also transferred into A. caldus MTH-04 with the help of plasmid RP4 or strains with plasmid RP4 integrated into their chromosome, such as E. coli SM10. The Kmr and Smr selectable markers from these plasmids were successfully expressed in A. caldus MTH-04. Futhermore, the IncP and IncQ plasmids were transferred back into E. coli cells from A. caldus MTH-04, thereby confirming the initial transfer of these plasmids from E. coli to A. caldus MTH-04. All the IncP and IncQ plasmids studied were stable in A. caldus MTH-04. Consequently, this development of a conjugational system for A. caldus MTH-04 will greatly facilitate its genetic study.
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KEYWORD
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Acidithiobacillus caldus, moderately thermophilic, conjugative transfer, IncP plasmid, IncQ plasmid
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