苏云金芽胞杆菌AiiA蛋白对魔芋软腐病菌的抗病活性

苏云金芽胞杆菌AiiA蛋白对魔芋软腐病菌的抗病活性 Activity of Bacillus thuringiensis AiiA Protein Against Soft Rot Disease for Amorphophallus konjac Caused by Erwinia.carotovora pv.carotovora 周燚 ZHOU Yi~ 1 2 3 first-author 孙明 1 2 3 喻子牛 1 2 3 华中农业大学生命科学技术学院农业微生物学国家重点实验室 华中农业大学生命科学技术学院农业微生物学国家重点实验室 1,2), SUN Ming~1, YU Zi-niu~1 1,2), SUN Ming~1, YU Zi-niu~1 华中农业大学生命科学技术学院农业微生物学国家重点实验室 湖北武汉430070长江大学农学院湖北荆州,434025 华中农业大学生命科学技术学院农业微生物学国家重点实验室 湖北武汉430070长江大学农学院湖北荆州,434025 ,湖北,武汉,430070 ,湖北,武汉,430070 AiiA蛋白通过破坏病原菌的信号分子来阻断病原菌的群体感应调节系统,达到抗病的目的,是一种新型的抗病蛋白.根据蜡状芽胞杆菌ATCC14579基因组中aiiA序列设计引物,从苏云金芽胞杆菌中成功扩增出aiiA基因,其ORF为753bp,测序后经blast检测与已发表的序列相似性均在95%以上.将此基因克隆到pET28a表达载体中,转化大肠杆菌BL21（DE3）,经IPTG诱导,超量表达的蛋白在菌体中以包涵体形式存在,纯化的AiiA蛋白经SDS PAGE电泳检测,表达产物分子量为28×103,致病性测定表明该蛋白对魔芋软腐病菌CZY具有较强的抗病活性. AiiA protein, blocking the bacterial quorum sensing by hydrolyzing AHL-lactone, can greatly attenuate the disease caused by many bacterial pathogens in which quorum sensing regulate the expression of virulence genes. The primers were designed from the sequence of aiiA gene in genome of B.cereus ATCC14579,and the 753 bp long gene segment was amplified from Bacillus thuringiensis by PCR （using） EX-Taq DNA polymerase. The comparability of aiiA gene from B.thuringiensis is very high with others. The PCR segment was inserted into expression vector pET28a（+）.Then the recombinant vector pETAiiA was introduced into E.coli BL21（DE3） for expression. SDS-PAGE showed that the expression product was a band of about 28×10 3 protein. The expression protein greatly attenuated the pathogenesis of E.carotovora pv.carotovora CZY which can result in soft rot for Amorphophallus konjac. 苏云金芽胞杆菌 AiiA蛋白 魔芋软腐病菌CZY Bacillus thuringiensis AiiA Protein Erwinia carotovora pv.carotovoraCZY S436.32 国家"973项目"（2003CB114201）;国家"863计划"资助项目（2004AA214092;2003AA223081）;国家自然科学基金资助项目（30270053）;教育部重点项目（01123） 2004-06-01 2021-04-01 6