抗砷载体的构建及在喜温硫杆菌中的接合转移  

Construction of the Arsenic Resistance Plasmids and the Conjugative Transfer to Thiobacillus caldus

作　　者：赵清[1] 刘相梅[1] 边疆[1] 詹杨[1] 刘缨[1] 林建群[1] 颜望明[1] 

机构地区：[1]山东大学微生物技术国家重点实验室,山东济南250100

出　　处：《生物技术》2005年第3期19-21,共3页Biotechnology

基　　金：国家重点基础研究项目(973)计划项目(2004CB619202);山东省优秀中青年科学家科研奖励基金计划(2004BS08006)

摘　　要：利用DNA体外重组技术,将质粒载体pUM3上的抗砷基因片段亚克隆到含有强启动子(tac启动子)并具有广泛寄主范围特性的IncQ族质粒pMMB24上,成功构建了含有强启动子的抗砷质粒pSDRA3,以及删除调节基因片段的组成型表达的抗砷质粒pSDRA4。通过接合转移的方式将其导入专性自养极端嗜酸性的喜温硫杆菌中,首次成功地建立了喜温硫杆菌的遗传转移系统,构建了冶金工程菌T.caldus(pSDRA3)和T.caldus(pSDRA4)。与野生菌相比,重组菌抗砷性能明显提高。A 4.3kb Hin d Ⅲ fragment harboring the arsenic resistance genes from plasmid pUM3 was subcloned into the wide-host-range IncQ plasmid pMMB24 and a new arsenic resistance plasmid pSDRA3 with tac promoter was constructed by using the recombinant technique in vitro.Then plasmid pSDRA4 was constructed by deleting the regulative gene,the lacI Q gene,from plasmid pSDRA3.Both plasmids pSDRA3 and pSDRA4 were transformed into E. coli JM109,and the arsenic resistance characteristics of the recombinant plasmids in E.coli were studied.Then,they were introduced into autotrophic extremely acidophilic Thiobacillus caldus by conjugative transfers. Compared with T.caldus ,the level of arsenic resistance of T.caldus (pSDRA3) and T.caldus (pSDRA4) were greatly raised.

关 键 词：抗砷基因 喜温硫杆菌 接合转移 

分 类 号：Q937[生物学—微生物学]