摘要
用抗性库蚊酯酶基因,引入原核表达载体pRL439,转化大肠杆菌HB101细胞,获得表达。通过酶切、Southern杂交鉴定重组质粒。研究了重组菌酯酶的活性,重组质粒pRLB1表达的酯酶具有高酶活并能高效降解酯酶的特异性底物α乙酸萘酯(αNA)和β乙酸萘酯(βNA);经对重组菌进行细胞固定化后降解农药三氯杀虫酯(7504),反应时间短。
Recombinant plasmid pRLB1 was constructed from detoxifying gene B1 of pesticide resistant \%Culex pipiens quinquefasciatus\% and from plasmid pRL439 contained the strong promoter PpsbA. The positive clone was identified by digestion and Southern analysis. Expression of recombinant plasmid containing esterase gene was detected. An engineered bacterium possessing high enzyme activity was obtained and immobilized. It can effectively degrade the specific substrate α\|naphthyl acetate (α\|NA) and β\|naphthyl of esterase enzyme. Assays showed that pesticide acetofenate (7504 an organic choride pesticide) was degraded by the immobilized cells within one hour.
出处
《微生物学报》
CAS
CSCD
北大核心
2000年第2期126-131,共6页
Acta Microbiologica Sinica
基金
国家自然科学基金! ( 396 80 0 0 1 )
山东省自然科学基金资助课题!(Y98D1 6 0 6 4 )
关键词
酯酶基因
基因表达
农药污染
水体净化
抗性库蚊
Esterase gene, Gene expression, \%Escherichia coli,\% Degradation pesticide
