苏云金杆菌毒性质粒在蜡状芽胞杆菌群间的水平转移  

Horizontal Transfer of BT Virulence Plasmid among Bacillus cereus sensu lato Population

作　　者：袁永明[1,2] 周帼萍[1,2] 刘海舟[1,2] 胡晓敏[1] 袁志明[1] 

机构地区：[1]中国科学院武汉病毒研究所,湖北武汉430071 [2]中国科学院研究生院,北京100049

出　　处：《微生物学杂志》2008年第2期1-5,共5页Journal of Microbiology

基　　金：国家自然科学基金项目(30470037)

摘　　要：以苏云金芽胞杆菌KT0为质粒供体菌,在液体环境中与蜡状芽胞杆菌组成员菌(Bacillus cereus sensu lato group strains)发生接合转移。对所筛选到的接合转移子进行了质粒稳定性检测、PCR验证以及供、受体菌染色体背景RAPD图谱分析。不同培养基中质粒转移率数据表明:在人工LB培养基及灭菌牛乳中,质粒pHT73均能以不同频率转移至蜡状芽胞杆菌组受体菌,并且在牛乳中转移活性更高,最高可这4.8×10^(-4)cfu/ donor。获得质粒的受体菌同时具备了质粒所编码基因功能,产生其编码的杀虫晶体蛋白。SDS-PAGE电泳和光镜结果显示转化菌在得到外源质粒pHT73后,能产生由cry1Ac基因编码的130 ku蛋白Cry1Ac,在芽胞期能形成稳定的菱形晶体。BT （Bacillus thuringiensis） subsp, kurstaki was taken as a KTO plasmid donor and synaptic transfer happened with Bacillus cereus sensu lato （BCSL） population in liquid conditions. The plasmid stability of the screened synaptic transferee was detected, and proved with PCR, and the chromosomal background of donor and receptor was analyzed with RAPD atlas. Data of plasmid transferring rate in different medium suggested that in artificial LB medium and sterile cow milk plasmid pHT73 could all transfer into BCSL receptor, and the transferring activity of those in cow milk was even higher and reaching 4.8 × 10^-4 cfu/donor at the highest. The plasmid receptor bacteria possesses at the same time functions coded by the plasmid, and produces insecticidal crystalline protein that coded. The results of SDS-PAGE electrophoresis and microscopy indicated that after obtaining exogenous plasmid pHT73 the transformant bacteria could produce 130 ku of Cry Ac protein coded by cryl Ac gene, and form stable rhombic crystal during the sporogenesis.

关 键 词：蜡状芽胞杆菌群 结合转移 pHT73 牛乳 

分 类 号：Q939.124[生物学—微生物学]