苏云金芽胞杆菌cry2Ad基因的克隆及其表达产物的活性分析  

Cloning,Expression and Insecticidal Activity of cry2Ad Gene from Bacillus thuringiensis

作　　者：张静涛[1,2] 束长龙[1] 宋福平[1] 刘楠[1] 张杰[1] 黄大昉[2] 

机构地区：[1]中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室,北京100193 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《生物技术通报》2009年第10期146-150,155,共6页Biotechnology Bulletin

基　　金："十一五"863计划(2006AA022189;2006AA10A212)

摘　　要：苏云金芽胞杆菌(Bacillus thuringiensis,Bt)SBT2是我国新分离出的一株野生菌株。扫描电镜显示该菌株产生双锥体形晶体。琼脂糖凝胶电泳发现其质粒图谱含有5个条带。聚丙烯酰胺凝胶电泳显示此菌株产生130kD晶体蛋白。利用PCR-RFLP法进行杀虫基因类型鉴定,发现其含有cry1Aa、cry1Da、cry1Hb、cry1Jb、cry1Ka、cry1Ib、基因。Cry2Ad蛋白的活性至今未见研究报道,本研究克隆和测序了该基因。并对其进行了表达。生物活性测定结果表明其表达产物对舞毒蛾(Lymant-ria dispar)、棉铃虫(Helicoverpa armigera)、亚洲玉米螟(Ostrinia furnacalis)、小菜蛾(Plutella xylostella)有低活性;对大猿叶甲(Colaphellus bowringi)无活性。A new Bacillus thuringiensis strain,SBT2,was isolated from soil samples in China. The strain formed the bipyramidal crystal observed by scanning electron microscopy. Five bands were found in the large plasmid profile of SBT2. SDS-PAGE analysis revealed molecular weight of insecticidal crystal protein in this strain was 130 kD. By use of PCR-RFLP identification method, cry1Aa, cry1Da,cry1Hb,cry1Jb,cry1Ka ,cryllb, cry2Ad genes were found in this isolate. The cry2Ad gene was cloned, sequenced and expressed. The bioassay results indicated that the expressed Cry2Ad had insecticidal activity against Lymantria dispar, Ostriniafurnacalis, Helicoverpa armigera and Plutella xylostella. But there was no activity against Colaphellus bowringi.

关 键 词：苏云金芽胞杆菌 cry2Ad基因 克隆 表达 生物活性 

分 类 号：S476[农业科学—农业昆虫与害虫防治]