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利用重叠延伸PCR技术,将苏云金芽胞杆菌中非芽胞特异性的cry3A基因的启动子替换cry1Ac10和cry1C基因的启动子序列,并用cry1Ac10 基因的终止子序列,替换cry1C 基因终止密码子下游的序列,得到改造的cry1Ac10和cry1C基因。改造的基因在苏云金芽胞杆菌无晶体突变株中可表达晶体蛋白,并形成伴胞晶体。这两个基因在转移到天然苏云金芽胞杆菌后可避免与其内源杀虫晶体蛋白基因竞争转录因子( 因子)从而提高杀虫晶体蛋白的表达量,为构建高效杀虫工程菌提供了有效的基因。
Using overlap extension PCR, the promoters of non-spore-specific cry3A gene in B. thuringiensis were substituted for the promoter sequences of cry1Ac10 and cry1C genes and the sequence of the stop codon of cry1Ac10 gene was replaced by the terminator sequence of cry1Ac10 gene to obtain Transformed cry1Ac10 and cry1C genes. The modified gene can express crystal protein in the Bacillus thuringiensis crystal-free mutant and form a companion crystal. The transfer of these two genes to natural Bacillus thuringiensis avoids competing transcription factor (factor) with their endogenous insecticidal crystal protein gene to increase the expression of insecticidal crystal protein, thus providing a valid gene for the construction of a highly efficient insecticidal engineering bacterium .