Harford N, Cabezon T, Crabeel M, Simoen E, Rutgers A, De Wilde M
Dev Biol Stand. 1983;54:125-30.
The genomes of HBV viruses of two different serotypes were cloned in E. coli. Sequences coding for the major polypeptides of surface antigen (HBsAg) were fused with the 5' end of a cloned yeast arg3 gene. When introduced into yeast, on a suitable vector, the hybrid gene directed the synthesis of a fusion protein. Crude extracts of such strains were shown to contain HBsAg like material having physical properties characteristic of the antigen isolated from the plasma of chronic human carriers, as judged by isopycnic and rate zonal centrifugation. Furthermore, these extracts readily elicit specific anti-HBsAg antibodies in rabbits. Further manipulations of the 5' part of the arg3 gene resulted in the introduction of a unique restriction site located in the 5' non translated leader sequence. The resulting vector was used to construct a recombinant plasmid directing the synthesis of the mature (226 amino acids) HBsAg polypeptide.
两种不同血清型的乙肝病毒基因组在大肠杆菌中被克隆。编码表面抗原（HBsAg）主要多肽的序列与克隆的酵母arg3基因的5'端融合。当通过合适的载体导入酵母时，杂交基因指导融合蛋白的合成。通过等密度和速率区带离心判断，这些菌株的粗提物显示含有具有从慢性人类携带者血浆中分离出的抗原特征物理性质的类似HBsAg的物质。此外，这些提取物很容易在兔子体内引发特异性抗HBsAg抗体。对arg3基因5'部分的进一步操作导致在5'非翻译前导序列中引入了一个独特的限制性酶切位点。所得载体用于构建指导成熟（226个氨基酸）HBsAg多肽合成的重组质粒。
