苏云金芽孢杆菌aiiA基因的克隆及植物表达载体构建

Cloning and construction of plant expression vector of aiiA gene from Bacillus thuringiensis

N-酰基高丝氨酸内酯(N-acyl-homoserine lactones,AHLs)是革兰氏阴性细菌致病过程中的关键信号分子,芽孢杆菌中广泛存在的aiiA基因编码AHL-Lactonase能够水解信号分子AHLs。利用PCR方法从苏云金芽孢杆菌中克隆了aiiA基因,序列分析表明该基因由753个碱基组成,编码含有250个氨基酸残基的蛋白质,核苷酸序列与已报道aiiA的同源性为89%～97%。将该基因连接到植物表达载体pCAMBIA1301中,成功构建了aiiA基因的植物表达载体pCAM-aiiA,为进一步通过转基因技术研究该基因的功能奠定了基础。

N-acyl-homoserine lactones （AHLs） is the crucial signal molecule when the gram-negative bacteria infects hose. The AHL-Laetonase expressed by aiiA gene which widespread in Bacillus thuringiensis, can hydrolyze AHLs.The study cloned the aiiA gene from Bacillus subtilis by PCR.The sequence analysis indicated that the clone was consisted of 751 nucleotides （nt）,which coding 250 amino acids. The nueleotide sequence showed 89%-97% identities with those of the aiiA gene that have reported. A plant expression vector of the aiiA gene was constructed. This work laid the foundations for future transgenic research on aliA gene function.