| 铜绿假单胞菌脂肪酶基因(LipA)在枯草芽孢杆菌pab02中的整合表达 |
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| 引用本文： | 张飞燕,何敏,王振华,潘康成,冯杰,唐慧琴.铜绿假单胞菌脂肪酶基因(LipA)在枯草芽孢杆菌pab02中的整合表达[J].湖南农业大学学报(自然科学版),2017,43(2). |
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| 作者姓名： | 张飞燕 何敏 王振华 潘康成 冯杰 唐慧琴 |
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| 作者单位： | 1. 四川农业大学动物医学院动物微生态研究中心,四川成都,611130;2. 四川农业大学动物医学院动物微生态研究中心,四川成都 611130;德阳市畜牧食品局,四川德阳618000;3. 成都农业科技职业学院,四川成都,611100 |
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| 基金项目： | "教育部长江学者和创新团队发展计划"资助项目，四川省高等学校科技创新团队资助项目 |
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| 摘 要： | 对铜绿假单胞菌脂肪酶基因(Lip A)进行克隆,构建整合表达质粒pHSG398–16S–lip A,并在枯草芽孢杆菌pab02中整合表达。根据铜绿假单胞菌Lip A基因设计引物,扩增Lip A基因,并进行测序,构建整合表达质粒pHSG398–16S–lip A,转化枯草芽孢杆菌pab02,并对转化后的阳性菌株进行诱导表达。结果显示,成功从铜绿假单胞菌A05菌株中扩增到长1 092 bp的Lip A基因,该基因与铜绿假单胞菌Lip A基因(登录号为AB290342)的同源性高达98%;该基因共编码311个氨基酸(aa),包含26个aa组成的信号肽和285个aa组成的成熟肽;成功构建了整合表达质粒pHSG398–16S–lip A,并将Lip A整合到枯草芽孢杆菌pab02染色体上,获得了重组枯草芽孢杆菌pab02–lip A;SDS–PAGE分析结果表明,脂肪酶蛋白得到了有效表达,该蛋白的相对分子质量约为37 000。
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| 关 键 词： | 铜绿假单胞菌 枯草芽孢杆菌 脂肪酶基因 |
Clone and integrative expression of gene lipase LipA from Pseudomonas aeruginosa in Bacillus subtilis |
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| ZHANG Feiyan,HE Min,WANG Zhenhua,PAN Kangcheng,FENG Jie,TANG Huiqin.Clone and integrative expression of gene lipase LipA from Pseudomonas aeruginosa in Bacillus subtilis[J].Journal of Hunan Agricultural University,2017,43(2). |
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| Authors: | ZHANG Feiyan HE Min WANG Zhenhua PAN Kangcheng FENG Jie TANG Huiqin |
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| Abstract: | This experiment aimed to construct an integrative expression vector pHSG398–16S–lipA, and expressed gene Pseudomonas aeruginosa lipase (LipA) in Bacillus subtilis pab02. A pair of primers was designed based on sequenceof gene LipA, and itsP. aeruginosa A05 was cloned and sequenced, the followed by the construction of integrative expression plasmid pHSG398–16S–lipA, then it was transformed intoBacillus subtilis pab02 for expression. The experiment obtained a 1092 bp length of LipA encoded with 311 amino acids, contained a signal peptide with 26 amino acids and a mature peptide with 285 amino acids, it shared 98% homology with gene lipase A of P. aeruginosa (Accession No. AB290342). The integrative expression plasmid pHSG398–16S–lipA was successfully constructed and transformed into B.subtilis pab02. Result from SDS–PAGE analysis showed that the lipase A was successfully expressed and its relative molecule weight was about 37000. |
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| Keywords: | Pseudomonas aeruginosa Bacillus subtilis lipase gene |
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