Kniskern P J, Hagopian A, Burke P, Dunn N, Emini E A, Miller W J, Yamazaki S, Ellis R W
Department of Virus and Cell Biology, Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania 19486.
Hepatology. 1988 Jan-Feb;8(1):82-7. doi: 10.1002/hep.1840080117.
The entire surface protein of hepatitis B virus serotype ayw containing the preS (preS1+preS2) and S domains has been expressed in the yeast Saccharomyces cerevisiae. Yeast containing a recombinant plasmid utilizing a constitutive promoter did not express this gene successfully due to the toxicity of the protein. A plasmid using a regulatable promoter directed expression which initiated late in the exponential phase of growth and resulted in the accumulation of high intracellular levels of the complete surface protein. The purified polypeptide aggregates into a form which, although not comprised of typical 20 nm particles, displays antigenic determinants encoded by the preS1, preS2 and S domains. Immunization of rabbits elicited the formation of antibodies directed against all three domains. This candidate vaccine will be useful for studying the contributions to viral immunity of the host response to the preS1 and preS2 domains.
乙型肝炎病毒ayw血清型包含前S（前S1 + 前S2）和S结构域的完整表面蛋白已在酿酒酵母中表达。含有利用组成型启动子的重组质粒的酵母由于该蛋白的毒性而未能成功表达该基因。使用可调节启动子的质粒指导的表达在生长指数期后期开始，并导致细胞内高水平完整表面蛋白的积累。纯化的多肽聚集成一种形式，尽管它不是由典型的20纳米颗粒组成，但展示了由前S1、前S2和S结构域编码的抗原决定簇。用其免疫兔子引发了针对所有三个结构域的抗体形成。这种候选疫苗将有助于研究宿主对前S1和前S2结构域的反应对病毒免疫的贡献。
