乙肝表面抗原在酿酒酵母中的异源表达

Heterologous Expression of HBsAg in Saccharomyces Cerevisiae

DOI：10.3969/j.issn.1673-1689.2015.09.002

中文关键词: 酿酒酵母 乙肝表面抗原 启动子 kozak序列 共表达

英文关键词: Saccharomyces cerevisiae, HBsAg, promoter, kozak sequence, co-expression

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中文摘要:

乙肝表面抗原HBsAg是乙肝疫苗的主要组分。从转录水平、翻译水平以及翻译后水平对其进行优化，实现了HBsAg在重组酿酒酵母中的高效异源表达。与组成型启动子TEF1、TDH3、HXT7、ADH1相比，诱导型启动子GAL1可大幅度提高HBsAg的表达；与GCCACC和AAAAAA相比，kozak序列TACACA最有利于HBsAg的翻译表达；而共表达二硫键异构酶pdi对HBsAg的活性表达具有显著的促进作用。通过3种水平的优化，HBsAg活性从最初的8.87 IU/g提高至151.08 IU/g；最终，利用Ni柱亲和层析成功获得纯化的HBsAg，活性为3.32 IU/mL。

英文摘要:

Hepatitis B surface antigen(HBsAg) is a major component of the hepatitis B vaccine. In the present work，we achieved its high efficient heterologous expression in the recombinant Saccharomyces cerevisiae by optimization at three levels:transcription，translation and post-translation. Compared with the constitutive promoters TEF1，TDH3，HXT7 and ADH1，the inducible promoter GAL1 substantially increased the expression of HBsAg. Moreover，compared with GCCACC and AAAAAA，the kozak sequence TACACA is more beneficial to the translation and active expression of HBsAg. More importantly，co-expression of protein disulfide isomerase(PDI) also significantly increased the expression of HBsAg. Through three levels of optimizations，the activity of recombinant HBsAg was increased from 8.87 IU/g CDW to 151.08 IU/g CDW. Finally，applying the Ni column affinity chromatography，we successfully obtained the purified HBsAg with an activity of 3.32 IU/mL.

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