Chen Si-Yong, Guo Min-Zhuo, Qiu Feng, Fei Yong-Liang, Yi Yao, Guo Yu, Jia Zhi-Yuan, Yu Tao, Bi Sheng-Li
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Feb;24(1):30-2.
To construct virus-like particles of hepatitis B core antigen, with HBsAg "a" epitope exposed on the surface.
Hepatitis B surface antigen "a" epitope were inserted into the Hepatitis B core antigen, between the 78th (Asp) and the 79th (Pro) amino acids. The gene was synthesized after the codon optimized, then it was ligated to the express vector after been enzyme digest. The virus-like particles were observed by electron microscope and detected by ELISA after been expressed and purified. Immune the rabbits by the VLPs, then detect the antibody.
The virus-like particles were confirmed by electron microscope. Its antigenicity and immunogenicity were identified by ELISA.
The prokaryotic express plasmid with the fusion gene has been constructed successfully. The virus-like particles have been expressed, purified and identified, which lays the foundation for its application in the further.
构建乙型肝炎核心抗原病毒样颗粒，使乙肝表面抗原“a”表位暴露于表面。
将乙型肝炎表面抗原“a”表位插入乙型肝炎核心抗原第78位（天冬氨酸）和第79位（脯氨酸）氨基酸之间。密码子优化后合成该基因，酶切后连接到表达载体。表达并纯化后，通过电子显微镜观察病毒样颗粒，并用酶联免疫吸附测定法进行检测。用病毒样颗粒免疫兔子，然后检测抗体。
通过电子显微镜确认了病毒样颗粒。通过酶联免疫吸附测定法鉴定了其抗原性和免疫原性。
成功构建了带有融合基因的原核表达质粒。病毒样颗粒已表达、纯化和鉴定，为其进一步应用奠定了基础。
