摘要
用改进的碱解法分析基因供体菌苏云金芽孢杆菌蜡螟变种81-6(Bacillus thuringiensisvar.galleriae 81—6)株有两种小质粒和一种大质粒。将碱解法抽提到的质粒用 PstI 完全酶解,与同样用 PstI 完全酶解的载体质粒 pCAMPUC 连接,以此重组质粒转化苏云金杆菌无晶体蛋白突变株 Bacillus thuringiensis var.kurstaki HD-1 Cry-B 株的原生质体。用ELISA 法筛选到一株阳性转化菌落,分析了其质粒组成和插入片段的大小。毒性试验显示阳性转化子对昆虫幼虫有毒杀作用。
Two small plasmids and one large plas-mid from Bacillus thuringiensis var.galleriaestrain 81-6 were detected by a simple,rapidmethod.The donor plasmid which was ex-tracted by alkaline method and the vectorplasmid pCAMPUC were digested with Pstland ligated with T4 ligase.The recombina-tion plasmid was transferred into protoplastsof Bacillus thuringiensis var.kurstaki HD-1cry-B strain.Transformants were screenedby ELISA method for the producing of crystalprotein in cry-B strain.A positive strain.was obtained.The insertion fragment ofcrystal protein gene was about 7.27 kb.Thebioassay of positive transformant showed toxiceffect to lepidopteran.
出处
《微生物学报》
CAS
CSCD
北大核心
1990年第4期249-253,共5页
Acta Microbiologica Sinica
关键词
苏云金杆菌
蜡螟变种
晶体蛋白基因
Bacillus thuringiensis
parasporal crystal protein gene cloning
