人工合成草鱼生长激素cDNA在大肠杆菌中的表达

经密码子优化的人工合成草鱼生长激素cDNA与表达载体pET-28a( )重组，构建重组表达质粒PET－GH。转化在肠杆菌BL21（DE3），筛选阳性克隆，IPTG诱导表达，12．5％的SDS－PAGE分析显示，大肠杆菌表达产物中含有与草鱼生长激素分子量一致的新增蛋白带，激光密度扫描，其产量约占菌体总蛋白的40％，金属离子螯合层析柱亲和纯化，获得电泳纯的重组蛋白。Western-blotting和酶联免疫吸附受体法检测证实，重组蛋白与抗草鱼生长激素的多克隆抗体发生特异性结合；复性合的重组蛋白有与天然草鱼生长激素一致的生物学活性。

Prokaryotic Expression of Recombinant Grass Carp Growth Hormone

The recombinant plasmid containing a synthetic grass carp growth hormone cDNA was transformed into Escherichia coli BL21 (DE3) and expressed efficiently,induced by lmmol/L IPTG. 12.5% SDS-PAGE showed the molecular weight of new added protein in Escherichia coli BL21 (DE3) is consistent with that of grass carp growth hormone. The recombinant protein was composed of approximately 40% of the total bacterial proteins. The target protein was purified with Ni-NTA affinity column.Western-blotting conformed that recombinant protein could specifically react with antibody against grass carp growth hormone. ELISA-receptor assay showed the bioacfivity of recombinant protein is consistent with that of nature grass carp growth hormone