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目的构建IFN α/HBVPreS2融合基因表达载体并进行表达。方法采用PCR技术分别扩增IFN α2b和HBVPreS2编码基因片段 ,并经分步克隆获得IFN α/HBVPreS2融合基因。然后将其插入质粒 pBV220 ,构建重组表达载体pBV IFN PreS2。结果经转化E.coli后 ,诱导表达出相对分子质量(Mr)为27000的融合蛋白。SDS PAGE分析显示 ,表达量占菌体总蛋白质的15 %。Westemblot分析表明 ,表达蛋白能与抗IFN α2b抗体结合。结论IFN α/HBVPreS2融合基因表达载体的构建 ,为进一步探讨HBV感染的特异性免疫治疗提供了实验依据。
Objective To construct IFNα / HBVPreS2 fusion gene expression vector and express it. Methods IFNα2b and HBVPreS2 gene fragments were amplified by polymerase chain reaction (PCR) and IFNα / HBVPreS2 fusion gene was obtained by stepwise cloning. Then, it was inserted into plasmid pBV220 to construct recombinant expression vector pBV IFN PreS2. Results After transformed into E. coli, a fusion protein with relative molecular mass (Mr) of 27000 was induced. SDS PAGE analysis showed that the amount of expression accounted for 15% of the total bacterial protein. Westemblot analysis showed that the expressed protein could bind to anti-IFNα2b antibody. Conclusion The construction of IFNα / HBVPreS2 fusion gene expression vector provides an experimental basis for further exploring the specific immunotherapy of HBV infection.