关键词: 抗虫蛋白, cryNGc基因, 原核表达, 等同性, 转基因

Abstract: The important precondition of safety evaluation of protein in genetically engineered maize is protein identification and equivalence study of proteins expressed in genetically engineered maize and Escherichia coli. CryNGc protein was expressed in E. coli through optimized methods such as codon optimization, cryNGc insecticidal protein gene synthesis, prokaryotic expression vector construction and E. coli expression conditions optimizition. On this basis, the characteristics and consistency of the recombinant protein HiII-NGc-1 in maize and CryNGc expressed in E. coli were analyzed by SDS-PAGE, immunoblot analysis and liquid chromatography-tandem mass spectrometry. The results showed that there was consistent in the expression of cryNGc gene in transgenic maize and E. coli by electrophoresis mobility, immune competence and amino acid composition consistent. The results confirmed that the CryNGc protein characteristics of maize transformants HiII-NGc-1 was consistent with the theoretical prediction, and was substantially equivalent to the CryNGc protein expressed in E. coli. The results provided the theoretical basis and data support for further biosafety evaluation of the new insect-resistant recombinant protein CryNGc maize transformants.

Key words: insect-resistant protein, cryNGc gene, prokaryotic expression, equivalence, transgenic maize