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含花生根瘤菌吸氢基因重组质粒pZ-55的特性分析

王建锋, 张凤章, 龙敏南, 王刚, 许良树

厦门大学生物学系.福建,厦门 361005

摘要

应用含豌豆根瘤菌部分吸氢基因的探针及PCR分析,从花生根瘤菌基因文库中筛选到含有吸氢基因的重组质粒pZ-55。用BamHⅠ、EcoRⅠ和KpnⅠ等内切酶对pZ-55进行酶切分析,构建了pZ-55的酶切物理图谱。经三亲本杂交分析,pZ-55能互补诱变株Ln-1(Hup~(-)),获得在自生条件下表达吸氢活性。

关键词： 花生根瘤菌; hup基因; 重组质粒pZ-55

Characterization analysis of recombinant plasmid pZ-55 containing hup gene from Rhizobium arachis

Wang Jianfeng, Zhang Fengzhang, Long Mingang, Wang Gang, Xu Liangshu

Department of Biology,Xiamen University.Xiamen 361005,Fujian

Abstract

One recombinant cosmid of pZ-55 was isolated from 71 colonies of the gene library by PCR screening and Dig-probe hybridization, which possibly contained hup genes. The size of insert foreign fragment in pZ-55 was about 19.6 kb. BamHI, EcoRI and KpnI were used in mapping analysis to indentify the physical organization of the insert foreign fragment. The cosmid of pZ-55 was transferred from E. coli strains to the Hup~(-) recipients: Ln-1 (a mutant of R. arachis derived from strain L8-3) by triparental mating using the plasmid pRK2013 as the helper plasmid. According to the analysis of plasmid DNA, the hybrid cosmids had been transconjugated into the recipients. The transconjugant EL-55 showed high level of H_(2) uptake activity when induced under free-living microaerobic condictions.

Key words: Rhizobium arachis;Recombinant plasmid pZ 55;hup gene

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引用本文：

王建锋, 张凤章, 龙敏南, 王刚, 许良树. 含花生根瘤菌吸氢基因重组质粒pZ-55的特性分析[J]. 生理学报 2000; 52 (1): .

Wang Jianfeng, Zhang Fengzhang, Long Mingang, Wang Gang, Xu Liangshu. Characterization analysis of recombinant plasmid pZ-55 containing hup gene from Rhizobium arachis. Acta Physiol Sin 2000; 52 (1): (in Chinese with English abstract).