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天然高产脂肪酶细菌的分离鉴定及脂肪酶基因的分析
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【中医学免费论文下载】作者:张迎光,施晓聪,林全思,王震,吕建新【摘要】 目的 :从环境中分离高产脂肪酶的细菌,并克隆其脂肪酶基因。方法:从天然高油脂含量土壤筛选高产脂肪酶的细菌,鉴定菌种后,设计相应引物PCR扩增脂肪酶基因并进行TA克隆,重组载体转入E.coli DH5α构建工程菌,双酶切及测序鉴定脂肪酶基因。结果:经过微生物菌种鉴定获得4株高活力菌,其中一株金葡菌产酶最高,利用PCR技术成功扩增脂肪酶基因,该基因经过克隆测序及其双酶切鉴定证实为新发现的突变脂肪酶基因,其二级结构与标准基因(EU310372)有较大不同。结论:成功分离一株高产脂肪酶的菌株——金黄色葡萄球菌,克隆的脂肪酶基因为新发现的突变型。 【关键词】 脂肪酶基因;PCR;TA克隆 Abstract: Objective: To separate natural bacteria producing high activities lipase from environment,and clone its lipase gene. Methods: High activities lipase producing bacteria was screened from soil which contained plenty of oils and fats. After identifying bacteria,PCR amplified lipase gene which was cloned to pMD18-T Vector. The recombint plasmid was transformed into E. Coli DH5αto build the engineering bacteria. At last,the lipase gene was identified with two restriction endonuclease digesting and sequencing. Results: After identifiy
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