植物病原菌群体猝灭基因克隆及诱导表达载体构建

摘要

N-acyl-homoserine lactones( AHLs) is crucial signal molecule when the gram-negative bacteria infects hose. The AHL-Lactonase,expressed by aiiA gene which widespread in Bacillus sp,can hydrolyze AHLs. The study have cloned the aiiA gene from Bacillus subtilis by PCR. The sequence analysis indicated that the clone was consisted of 751 nucleotides( nt) , coding 250 amino acids. The nucleotide sequence showed 87% -96% identities with those of the aiiA gene that have reported. A plant expression vector of the aiiA gene was constructed and named Pcamb-PPP3-aii4 ,in which the aiiA was controlled by the PPP3 promoter. This work laid the foundations for future transgenic research on aiiA gene function.%N-酰基高丝氨酸内酯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A HL-Lactonase能够水解信号分子AHLs.研究利用PCR方法从枯草芽孢杆菌中克隆了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87％ ～ 96％.将该基因置于诱导型启动子PPP3调控下,连接到植物表达载体pCAMBIA1301中,成功构建了aiiA基因的植物诱导表达载体pCAM -PPP3 -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.