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对铜绿假单胞菌脂肪酶基因(LipA)进行克隆,构建整合表达质粒pHSG398–16S–lipA,并在枯草芽孢杆菌pab02中整合表达.根据铜绿假单胞菌LipA基因设计引物,扩增LipA基因,并进行测序,构建整合表达质粒pHSG398–16S–lipA,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1092 bp的LipA基因,该基因与铜绿假单胞菌LipA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–lipA,并将LipA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–lipA;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37000.“,”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.