Construction of a genetically engineered and stable strain of degrading gamma-hexachlorocyclohexane and carbendazim by transposon mini-Tn5.
The complete dehydrochlorinase gene linA of a gamma-hexachlorocyclohexane (gamma-HCH) degrading strain Sphingomonas sp. BHC-A, containing promoter and Shine-Dalgarno sequence (SD sequence), was amplified by PCR. The linA gene was inserted into NotI-cut transposon vector pUT/mini-Tn5 (Km(r)) to get a novel transposon vector pUT/mini-Tn5-linA. With the helper plasmid RK600, the transposon vector pUT/mini-Tn5-linA was introduced into one carbendazim degrading gram-positive strain Rhodococcus sp. DJL-6 by triparental conjugation and then the dehydrochlorinase gene linA was integrated into the chromosome of Rhodococcus sp. DJL-6 by the transposon mini-Tn5. The selected multifunctional genetically engineered strain DJL-6A could degrade gamma-HCH and carbendazim simultaneously. The dehydrochlorinase activity of DJL-6A was as strong as that of Sphingomonas sp. BHC-A in 0.05 and 5 microg/mL initial gamma-HCH concentration. The linA of the strain DJL-6A was genetically stabile after successive plating DJL-6A for 30 days on nonselective media.
Wu
Jun
J
Key Lab of Microbiological Engineering Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China. wujunchina2002@yahoo.com.cn
Xu
Jingliang
J
Hong
Qing
Q
Li
Shunpeng
S
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001562:Benzimidazoles / Q000378:metabolism*; D002219:Carbamates / Q000378:metabolism*; D004251:DNA Transposable Elements; D005659:Fungicides, Industrial / Q000378:metabolism*; D005818:Genetic Engineering; D001556:Hexachlorocyclohexane / Q000378:metabolism*; D007306:Insecticides / Q000378:metabolism*; D008190:Lyases / Q000235:genetics / Q000378:metabolism; D016254:Mutagenesis, Insertional; D012240:Rhodococcus / Q000235:genetics* / Q000378:metabolism; D020578:Sphingomonas / Q000201:enzymology
D001426:Bacterial Proteins; D001562:Benzimidazoles; D002219:Carbamates; D004251:DNA Transposable Elements; D005659:Fungicides, Industrial; D007306:Insecticides; D001556:Hexachlorocyclohexane; D008190:Lyases; C019994:dehydrochlorinases; C006698:carbendazim