Cupriavidus metallidurans (C.metallidurans) CH34 is a kind of heavy metalresistant bacterium, whose genome own two phenoldegrading gene clusters and can grow on the medium that has phenol, cresol, benzoic acid, aniline as sole carbon and energy source. By using vector pIndigoBAC 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 phenoldegrading 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.
LI Li,ZHANG Jun-ya,TANG Yuan-ping,SONG Ren-tao,ZHU Chen-guang
. Screening and Characterization of Two PhenolDegrading 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