[含两个多克隆位点的乙肝病毒包膜蛋白与核心蛋白融合表达载体的构建及表达]
[Construction and expression of hepatitis B virus envelope protein combined with core protein with two multiple cloning sites vector].
作者信息
Shen Hong-Hui, Liu Su-Jun, Guo Lei, Hou Jun, Wang Zhi-Jie, Xin Shao-Jije, Bai Bing-Ke, Hu Yan, Yu Xiang-Hui, Kong Wei, Mao Pan-Yong
机构信息
College of Life sciences, Jilin University, Changchun 130012, China.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Aug;23(4):313-5.
OBJECTIVE
To develop a coexpression plasmid which expressing envelope protein and nucleoprotein of hepatitis B virus and know of its expressing efficiency.
METHODS
The plasmid coexpressing envelope protein and nucleoprotein of hepatitis B virus under the CMV promoter respectively was constructed by gene recombination. Cellular expression was assessed by ELISA.
RESULTS
Multiple cloning site was inserted into expression vector contain hepatitis B virus PreS2-S gene. And expression unit containing hepatitis B virus PreC-C was cloned into it. HBsAg and HBeAg was detected both in the culture supernatant and in the cells.
CONCLUSION
The coexpressing plasmid was constructed successfully and it can express effectively in vitro. This has provided a basis for further research of the therapeutic HBV DNA vaccine.
目的
构建一种共表达乙型肝炎病毒包膜蛋白和核蛋白的质粒，并了解其表达效率。
方法
通过基因重组构建分别在巨细胞病毒启动子下共表达乙型肝炎病毒包膜蛋白和核蛋白的质粒。采用酶联免疫吸附测定法评估细胞表达情况。
结果
将多克隆位点插入含有乙型肝炎病毒前S2-S基因的表达载体中。并将含有乙型肝炎病毒前C-C的表达单元克隆到其中。在培养上清液和细胞中均检测到乙型肝炎表面抗原和乙型肝炎e抗原。
结论
成功构建了共表达质粒，且其能在体外有效表达。这为进一步研究治疗性乙肝DNA疫苗提供了依据。
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