新型生长抑素原核表达质粒的构建及表达鉴定

Construction and Characterization of a Novel Somatostatin Prokaryotic Expression

摘要:

将生长抑素(SS)与乙肝表面抗原(S)融合基因插入平衡致死系统原核表达质粒pYA3493中, 转化至缺失asd基因的减毒猪霍乱沙门氏菌C500, 经酶切、测序筛选得到非抗性的目的克隆, 命名为pYA-SS。应用SDS-PAGE和Western blotting 技术分离并检测融合蛋白在宿主菌中的表达活性。结果表明, 本试验构建的非抗性筛选生长抑素原核表达质粒可以在宿主菌C500中稳定、正确表达。此研究为开发新型、高效、安全的促生长疫苗提供了可靠的研究材料。

Abstract:

In the current work, the fusion gene including somatostatin (SS) and the hepatitis B surface antigen gene was cloned into a balanced lethal system plasmid (pYA3493), and then transformed into asd? attenuated Salmonella choleraesuis C500 strain, the positive transformant without antibiotic resistance gene was confirmed by restriction analysis and DNA sequencing, designated as pYA-SS. The expression and immunogenicity of fusion protein were detected by SDS-PAGE and Western blot analysis. These results show that the recombinant prokaryotic expression plasmid pYA-SS could express the SS fusion protein with good immunogenicity in C500 strain. In above all, this study could provide reliable materials to develop novel, good and safe vaccine in enhancing the growth of animals.