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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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Citing This Article：

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).