苏云金芽胞杆菌cryI基因探针的制备及质粒基因定位  

Preparation of Bacillus thuringiensis cryI RNA Probe and Use of the Probe for Plasmid Gene Location

作　　者：关雄[1] 黄志鹏[1] 高日霞[1] 

机构地区：[1]福建农业大学植物保护系

出　　处：《福建农业大学学报》1996年第3期329-333,共5页Journal of Fujian Agricultural University

基　　金：国家计委85-504-21-01攻关课题;福建省教委资助项目

摘　　要：将ｐＥＳＩ经ＥｃｏＲＩ酶解后回收纯化的含有苏云金芽胞杆菌ｃｒｙＩ基因高保守区的ＥｃｏＲＩ－Ｆ片段重组到ｐＳＥＬＥＣＴ－１的ＥｃｏＲＩ位点，通过ＤＩＧ体外标记转录制备高灵敏度、背景清楚的ＲＮＡ探针，并利用该探针进行苏云金芽胞杆菌８０１０杀虫基因定位．结果表明。The EcoRI F fragment, which harbors highly conserved domain of Bacillus thuringiensis insecticidal crystal protein cryIA gene, was obtained through pESI digested with EcoRI. This fragment was then recovered with Glassmilk protocol and was ligated into EcoRI site of pSELECT 1 vector. The RNA probe was prepared by DIG labelled transcription system in vitro. It had higher sensitivity and lower background than DNA probe. At the same time, the gene was located by using this probe. The results provided positive evidence that the gene encoding the insecticidal crystal protein of Bt 8010 was located on the plasmid of 37×10 6.

关 键 词：苏云金芽胞杆菌 cryI基因 RNA探针 基因定位 

分 类 号：Q78[生物学—分子生物学]