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将一段人工合成的编码HBsAg 226个氨基酸的基因(adw亚型)克隆到酵母表达载体pGPD中,构建成pGPD-1(HBs)。HBsAg基因的5′未端被一种化学合成的DNA片段所取代,此片段在含有表达载体pGPD-1(HBs)的啤酒酵母株RH218中最易转译起始。采用高细胞密度发酵法发酵,HBsAg的产量与细胞浓度呈线性增加。HBsAg的最终浓度是54mg/L,占整个细胞蛋白质重量的0.14%。作者观察了酵母中HBsAg颗粒形成的机理,发现在细胞溶解产物中HBsAg的浓度取决于细胞溶解和抽提制备的方法。HBsAg颗粒未被糖基化而是以一种脂蛋白
A synthetic 226 bp gene encoding HBsAg (adw subtype) was cloned into the yeast expression vector pGPD to construct pGPD-1 (HBs). The 5 ’end of the HBsAg gene was replaced by a chemically synthesized DNA fragment, which was most easily initiated by translation in the Saccharomyces cerevisiae strain RH218 containing the expression vector pGPD-1 (HBs). Using high cell density fermentation fermentation, HBsAg production and cell concentration showed a linear increase. The final concentration of HBsAg is 54 mg / L, accounting for 0.14% of the total cellular protein weight. The authors looked at the mechanism of HBsAg particle formation in yeast and found that the concentration of HBsAg in cell lysates depends on the method of cell lysis and extraction preparation. HBsAg particles are not glycosylated but rather as a lipoprotein