Acquire Of HCH Degrading Enrichment,Cloning Of Genes Involved In HCH Degrading And Exoressing Of Dehydrochlorinase Gene

Environmental pollution is a worldwide problem. Bioremediation is a good way and has more merits. So it becomes the focus of researchers, but there are some questions before the isolation of degrading bacteria. It is hard to find them from enrichment and some of them have no competitive ability in our environment. The genes cloned from pure culture are single for the source of gene is single. The degrading ability of enrichment including all kinds microbacteria are high effective for their cooperation action. So how to exploit the enrichment is a question worth being study and discussion. In this article, some research was done about the enrichment and try to find new way to study the degrading of toxic compound.The soil polluted by HCH was collected and enriched in the laboratory. Four isomers of HCH(5vppm) could be degraded by enrichment in 36h at 30℃ with and 5% inoculation. No pure culture was got from the enrichment, so some study was done to exploit the enrichment for use in bioremediation. Study shows the enrichment could be amplified as pure culture. The time of formation of enrichment by seeding tank and fermenter are 28h and 20h under conditions of 30℃,300 r/min rotation,with 5% and 10% inoculation, 6L/min and 60L/min aeration.The total DNA from polluted soil and enrichment was extracted and purified to adapt the study of molecular biology. The total DNA was used as template for PCR amplification with conserved lin genes primer. Five genes involved in the degrading of HCH was cloned from total DNA of enrichment, such as linN, linB1, linC1, linD1, linE1, and sequenced. They all shows more than 98% homogenous to the genes got from pure culture in published papers. The G+C content of the linN gene is considerably lower than that of the other genes and the genomic DNA of the S. paucimobilis strains, suggesting that the linN gene may have originated from another organism with a lower G+C content, this may be the result of the gene level transfer. There are at least two pure culture in...