Harford N, Cabezon T, Colau B, Delisse A M, Rutgers T, De Wilde M
Department of Molecular and Cellular Biology, Smithkline Biologicals, Rixensart, Belgium.
Postgrad Med J. 1987;63 Suppl 2:65-70.
A host/vector system suitable for large-scale production of HBsAg has been constructed and optimized in terms of the expression plasmid and yeast host strain in order to permit fermentation to very high cell densities. The final expression plasmid contains the coding sequence of the major HBsAg protein (P24) flanked by the promoter sequences from a glycolytic gene and by the transcription-termination region of the ARG3 gene. The host/vector system was found to be genetically stable under large-scale fermentation conditions as demonstrated by nucleotide sequencing and restriction mapping experiments. The P24 protein is recovered from yeast as particles whose physiochemical properties are very similar to those of plasma-derived HBsAg.
为了实现极高细胞密度的发酵，已构建并优化了一种适用于大规模生产乙肝表面抗原（HBsAg）的宿主/载体系统，涉及表达质粒和酵母宿主菌株。最终的表达质粒包含主要乙肝表面抗原蛋白（P24）的编码序列，其两侧分别是来自糖酵解基因的启动子序列和ARG3基因的转录终止区域。通过核苷酸测序和限制性图谱实验证明，该宿主/载体系统在大规模发酵条件下具有遗传稳定性。P24蛋白从酵母中回收时呈颗粒状，其物理化学性质与血浆来源的乙肝表面抗原非常相似。
