【摘要】 酵母SUC2基因克隆YFD6的DNA用核酸酶BAL31从SUC2基因的BamHI切点进行酶解,加上BamHI接头后,自连环化,得到的一个缺失质粒 YFD6△21保留 SUC2基因的启动子-信号顺序以及氨基端22个氨基酸残基的编码顺序。将带有 HBsAg 基因的 BamHI 片段插入 YFD6△21的 BamHI 切点,使 HBsAg 基因和 SUC2基因融合,然后将重组质粒引入酵母。将酵母转化子在葡萄糖阻遏及去阻遏条件下生长,然后测定细胞内、周质空间和培养基中的 HBsAg 的量。我们只能在葡萄糖去阻遏条件下检测到 HBsAg 的表达,几乎全部表达的HBsAg 位于细胞内。更多还原

【Abstract】 Plasmid DNA YFD6,the yeast SUC2 gene clone,was digested with nuclease BAL31 from BamHI site of SUC2 gene.After adding BamHI linker,DNA moleculars were ligated by T4 DNA ligase.One of deletion plasmid,YFD6△21,remains SUC2 promoter-signal sequence and the sequencecoding for N-terminal 22 amino acid residues of external invertase.InsertBamHI fragment which contains HBsAg gene in BamHI site of YFD6A21and make gene fusion between SUC2 gene and HBsAg gene.Then transformrecombinant plasmids into yeast.rowing resulting transformants at glucose repressed or derepressed condition,the level of HBsAg expressed in intracellular part,periplasmic spaceand medium was determinated.We could detect expression of HBsAg geneonly at glucose derepressed condition.Almost all of expressed HBsAg wasin the intracellular part.更多还原