[Cloning, expression of the gene of dehydrochlorination of BHC from enrichment and its degradation experiment].
A high effective enrichment that could degrade four isomers of BHC completely was got from the soil polluted by gamma-BHC, but the pure culture was not obtained yet. The 471 bp sequence of linN was amplified from the total DNA of enrichment by polymerase chain reaction (PCR). The nucleotide sequence analysis showed that the linN gene had high homology with reported linA up to 99%. Then the amplified fragment linN was cloned in the proper orientation into the site between Nde I and Hind III of pET-29alpha via restriction endonuclease Nde I and Hind III. The recombinant was transformed into its host E. coli strain BL21 and a recombinant protein of about 17 kD was highly expressed and showed high ability of degrading gamma-BHC after induced by IPTG. The expressed protein occupied about 30% of the total bacterial protein. The cell extracts also showed some ability of degradation of gamma-BHC. It offered basic theory for the isolation of pure culture and the construction of genetic engineering microorganisms.
Zhang
Guo-shun
GS
Key Lab of Microbiological Engineering Agricultural Environment, Ministry of Agriculture, College of Life Science, Nanjing Agricultural University, Nanjing 210095, China. sheyangzhang@sohu.com
Hong
Qing
Q
Ma
Ai-zhi
AZ
Zhang
Xiao-zhou
XZ
Li
Shun-peng
SP
D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D001556:Hexachlorocyclohexane / Q000378:metabolism*; D007306:Insecticides / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012689:Sequence Homology, Nucleic Acid; D012988:Soil Microbiology; D020578:Sphingomonas / Q000235:genetics* / Q000378:metabolism
D007306:Insecticides; D011994:Recombinant Proteins; D001556:Hexachlorocyclohexane