Cloning Of The Gene Of Degratating Synthetic Pyrethroid Pesticide

The Acinetobacter sp. JXCC2D can degrade synthetic pyrethroid pesticide and other pesticides efficiently and broadly. The genomic library of the bacteria was constructed successfully, with the E.coli JM110 as the receptor strain, and pGEM-3Z as the vector. Both strain and that containing the vector could hardly degrade the pesticides which were tested in this paper. Partly digested the Acinetobacter sp.JXCC2D genomic DNA ,with MobI, into the fragments between 3kb and 7kb. Completely digested the pGEM-3Z, and purified the linear as the vector. The two restriction endonucleases are isocaudamer, and have the same coherent terminal. We got the genomic library with the insert fragments among 2-7kb, and the capacity was 10~4.The white single colony was incubated on the Burk solid medium containing the pesticides, and the esterase screening solid medium, at 37℃. The colony which contains the target gene can grow in both plates, and it can make the transparent zone. One colony containing the degrading gene was isolated, named 2-4-38, whose excellar esterase relative activity was 0.535. The 2-4-38 was incubated with the 0.2g/L pesticides for 108 h. Its degradation ratio to b-cypermethrin, cyfluthrin and deltamethrin were 66.0%, ,50.5% and 95.7%, respectly. Its degradation radio to profenofos and pyridaben were 32.5% and 25.9%, respectly, when cultured for 72h.The p2-4-38 contains a 3.2kb insert fragment. Sequenced and analysised this fragment. It was concluded that there was a ORF composed by 617 amino acids, and it had 94% identity with the lipase of MIS38 (Pseudomonas sp.) . It was inferred that the degradation gene was a member of lipase. The sequence was submitted to the Genebank, and the accession number was DQ906143.There is no report that the lipase can degrade the pesticides. So this paper cloned the lipase gene which has the activity of pesticides-degradating at the first time.