人乳头瘤病毒多肽与人免疫球蛋白G融合表达

目的 将人乳头瘤病毒16型(Human papillomavirus type 16,HPV-16)的晚期表达蛋白E7上的抗原24肽(从第38位氨基酸到第61位氨基6病毒感染防治酸)与人免疫球蛋白G的重链恒定区融合表达,并以此融合蛋白作为抗原,可能为HPV-1提供免疫治疗方法。方法 利用PCR方法分别扩增HPV-16 E7(38-61)24肽的DNA片段和人免疫球蛋白G的重链恒定区DNA片段,并构建到pEV21a表达载体上,转化入E．coli中表达,利用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹(Western-blotting)的方法对表达结果进行鉴定。结果 构建的表达载体HPV16E7e／hIgGHCCR-pET21a经酶切鉴定和测序显示序列正确;通过SDS-PAGE和Western-blotting的鉴定,重组融合蛋白Mr约40000,表达量可占菌体蛋白的20％左右。结论 成功构建HPV16-E7的抗原多肽片段和人免疫球蛋白G的重链恒定区的融合蛋白,并可在E．coli中高效表达。

Expressions of HPV16 peptide fused to the fragment of human immunoglobulin G

Objective To express the 24 peptide of human papillomavirus type16 (HPV16) E7 fused to the heavy chain constant region of human immunoglobulin G (IgG) in order to develop a candidate vaccine against HPV 16. Methods The DNA fragments of HPV16 E7 24 peptide and human IgG heavy chain constant region were amplified by PCR, then insert into the pET21a to construct an expression plasmid HPV16E7e/hIgGHCCR-pET21a. The expression plasmid was expressed in E.coli and the expression was identified by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Results The expression plasmid HPV16E7e/hIgGHCCR-pET21a was confirmed by restriction digestion and sequencing. The molecular weight of the chimeric protein was about M r 40 000, 20% of all bacteria protein. Conclusion The chimeric protein is constructed, which can overcome the low expression of HPV16 E7 and be highly expressed in E.coli.