Life Science

Screening and Characterization of Two PhenolDegrading Gene Clusters from Cupriavidus metallidurans CH34

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  • (Shanghai Key Laboratory of Bio-energy Crops, School of Life Sciences, Shanghai University, Shanghai 200444, China)

Online published: 2012-08-30

Abstract

Cupriavidus metallidurans (C.metallidurans) CH34 is a kind of heavy metalresistant bacterium, whose genome own two phenoldegrading gene clusters and can grow on the medium that has phenol, cresol, benzoic acid, aniline as sole carbon and energy source. By using vector pIndigoBAC 5, a bacterial artificial chromosome (BAC) library of C.metallidurans CH34 was constructed and thirty thousand clones were obtained, from which 98% of the clones had an insertion fragment with average size of 30 kb. It was deduced that the whole BAC library size is about 1 240 times of the genome size. More than 3 000 clones were screened with PCR and nine positive clones were obtained: five with longer gene clusters and four with shorter clusters. Two clones with total length of phenoldegrading gene cluster were used to study its expression in Escherichia coli (E.coli) on the medium using phenol as the sole carbon source. The results indicate that both clones have the ability to degrade phenol, and the one carrying a shorter cluster shows stronger ability than that of the one with longer cluster.

Cite this article

LI Li,ZHANG Jun-ya,TANG Yuan-ping,SONG Ren-tao,ZHU Chen-guang . Screening and Characterization of Two PhenolDegrading Gene Clusters from Cupriavidus metallidurans CH34[J]. Journal of Shanghai University, 2012 , 18(4) : 435 -440 . DOI: 10.3969/j.issn.1007-2861.2012.04.020

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