新型生长抑素原核表达质粒构建与表达鉴定.PDFVIP

生 物 工 程 学 报 Chin J Biotech 2008, June 25; 24(6): 995-998 Chinese Journal of Biotechnology ISSN 1000-3061 cjb@ © 2008 Institute of Microbiology, CAS CSM, All rights reserved 研究报告 新型生长抑素原核表达质粒的构建及表达鉴定 梁爱心, 冯细钢, 韩丽, 滑国华, 桑雷, 刘兴斌, 刘耘, 杨利国 华中农业大学动物科技学院, 武汉 430070 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。 关键词: 生长抑素, 原核表达, 平衡致死系统 Construction and Characterization of a Novel Somatostatin Prokaryotic Expression Aixin Liang, Xigang Feng, Li Han, Guohua Hua, Lei Sang, Xingbin Liu, Yun Liu, and Liguo Yang College of Animal Science Technology, Huazhong Agricultural University, Wuhan 430070, China 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 SDSand 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