关键词: 双价载体;高效;抗虫性;表达分析

Abstract:

In this study, the Bt and cpti gene were modified on the base of existing vector and expression vector. Soybean trypsin inhibitor SKTI signal peptide sequence was added at 5′ end of cpti, KDEL signal peptide sequence was added at its 3′ end, and at Bt 5′ end chloroplast targeting peptides was added, and then a bivalent expression vector PGBIC(K).B.4A with signal peptides and a control vector PGBIC.B.4A without signal peptides were constructed. Through pollen tube pathway mediated by agro-bacterium Gossypium hirsutum Y18 was transformed. The result of southern blot showed that the exogenous gene had been integrated into the receptor with genome different copies. By ELISA, it was found that the protein expression was increase by 1~8 times in 5 positive strains. In the insecticidal tests, the modified binary vector transgenic plants1/4-1showed higher resistance to the bollworm because of the accumulation of protein. This study provided a new method to obtain the higher resistance of bivalent transgenic cotton, and basis to improve the accumulation of other foreign protein in genetic engineering.

Key words: bivalent vector, high-performance, insect resistant, expression analysis