摘要
苏云金芽孢杆菌变种Bacillus thuringiensis aizawai 7-29 δ-内毒素基因及其3′末端缺失基因,亚克隆到质粒pUC19上,构建了pUCF33和pUCK63重组质粒。进一步将δ-内毒素基因及其3′端缺失基因,插入到植物表达载体pBI121.2质粒中,构建了pGY61和pGYCK63重组质粒。用^(32)中标记的δ-内毒素3′末端缺失的Kpn I DNA片段的探针,与重组质粒进行DNA-DNA分子杂交,结果阳性。带有全长和3′末端缺失重组质粒,转入大肠杆菌HB101和农杆菌LA4404受体细胞中,其克隆子细胞抽提物,对大菜粉蝶和玉米螟幼虫均具有杀虫生物活性。结果证明δ-内毒素基因不仅能在大肠杆菌HB101中,而且能在农杆菌LA4404中表达。
The δ-endotoxin gene from Bacillus thuringiensis subsp. aizawai 7-29 and the δ-endotoxin gene with 3′ end deletion were subcloned intopUC19 plasmid, resulting recombinant plasmids pUCF33 and pUCK63,respectively. After amplification of recombinant plasmids, pGY61 andpGYCK63 were constructed by the insertion of δ-endotoxin gene andits end deletion into plant vector pBI121.2. The DNA of recombinantplasmids pGY61 and pGYCK63 were hybridized positively with α-^(32)P-labelled Kpn I DNA fragment of 3′end deletion of δ-endotoxin gene.The full length δ-endotoxin gene and δ-endotoxin gene with 3′ enddeletion were expressed in both E. coli HB101 and A. tumefaciensLA4404. The cell extracts from transformants of E. coli HB101 and A.tumefaciens LA4404 are lethal to Pieris rapae larvae and Ostrinia nubi-lalis Iarvae.
出处
《生物工程学报》
CAS
CSCD
北大核心
1991年第1期54-61,共8页
Chinese Journal of Biotechnology
关键词
δ内毒素基因
亚克隆
苏云金杆菌
δ-endotoxin
subcloning
recombinant
deletion
Bacillus thuringiensis
