摘要
对苏云金芽孢杆菌鲇泽变种(Bacillus thuringiensis var.atzawai)7-29杀虫蛋白质基因调控区(181bp)和第五活性区(217bp)修饰后,在它的5′端和3′端分别插入一个人工合成并带有启动密码子(ATG)的 adapter 1(15bp)和 PCR 合成并带有终止密码子(TAA)的第五活性区(229bp)。使负责编码 N-端肽的 DNA 片段,改造成既有启动翻译功能又有终止翻译功能的结构基因。经过 Western blotting 检测,改造后的结构基因能在 E.coli JM103中正常表达。生物杀虫定性实验结果表明,3′端和5′端全改造后的杀虫基因比只改造3端的杀虫基因更具有较好的杀虫活性。
The regulative region(181bp)and the fifth toxic active domain(217bp)were removedfrom the insecticidal protein gene of Bacillus thuringiensis var.aizawai 7—29.After the syn-thesis of the adaptor(15bp)that contains initiation codon(ATG)and the PCR synthesis ofthe fifth toxic active domain(229bp)that contains stop codon(TAA),were inserted into on5′truncated and 3′truncated of the coding fod N-terminal peptid's DNA fragment,that to be-come a modified structural gene.The modified structural gene can be play initiatic translation-function and stop translation-function during translation of insecticidal protein.The insecticidalprotein was determined by western blotting,showed the expression of modified structural genein Escherichia coli JM 103.The bioassay of insecticidal proteins showed the 3′truncated and5′truncated of insecticidal gene was higher toxic active than the 3′truncated of insecticidalgene in Escherichia coli JM 103.
出处
《微生物学报》
CAS
CSCD
北大核心
1992年第3期167-175,共9页
Acta Microbiologica Sinica
基金
本课题属“七五”攻关
非教育系统留学回国人员择优资助课题
关键词
鲇泽变种
杀虫蛋白基因
苏云金杆菌
B(?)cillus thuringiensis var.aizawai 7—29
Insecticidal gene
Modification
Expression
