Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Construction of Conjugative Gene Transfer System Between E. coli and Moderately Thermophilic, Extremely Acidophilic Acidithiobacillus caldus MTH-04

  • Liu, Xianggmei (State Key Laboratory of Microbial Technology, Shandong University) ;
  • Lin, Jianqun (State Key Laboratory of Microbial Technology, Shandong University) ;
  • Zhang, Zheng (State Key Laboratory of Microbial Technology, Shandong University) ;
  • Bian, Jiang (State Key Laboratory of Microbial Technology, Shandong University) ;
  • Zhao, Qing (State Key Laboratory of Microbial Technology, Shandong University) ;
  • Liu, Ying (State Key Laboratory of Microbial Technology, Shandong University) ;
  • Lin, Jianqiang (State Key Laboratory of Microbial Technology, Shandong University) ;
  • Yan, Wangming (State Key Laboratory of Microbial Technology, Shandong University)
  • Published : 2007.01.31
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Abstract

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 $Km^r\;and\;Sm^r$ 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.

Keywords

References

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