Expression of hepatitis B surface antigen in yeast.
Expression of hepatitis B surface antigen in yeast.
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发表时间：
1983
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影响因子：
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通讯作者：
N. Harford;T. Cabezón;M. Crabeel;E. Simoen;A. Rutgers;M. de Wilde
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作者：
N. Harford;T. Cabezón;M. Crabeel;E. Simoen;A. Rutgers;M. de Wilde
在E.杆菌将编码表面抗原（HBsAg）主要多肽的序列与克隆的酵母arg3基因的5'端融合。当在合适的载体上引入酵母时，杂合基因指导融合蛋白的合成。这些菌株的粗提物显示含有HBsAg样物质，其具有从慢性人携带者血浆中分离的抗原的物理特性，如通过等密度和速率区带离心所判断的。此外，这些提取物很容易在家兔中引发特异性抗HBsAg抗体。对arg3基因的5'部分的进一步操作导致引入位于5'非翻译前导序列中的独特限制性位点。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。
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.