中文摘要：

目的：利用分子克隆技术在原核细胞中研究14-3-3γ融合蛋白的表达及分离纯化。方法：采用RT—PCR法从大鼠心肌组织中扩增14-3—3γcDNA编码区，并将其重组于谷胱甘肽硫转移酶（GST）融合蛋白表达质粒pGEX—KG中，免疫印迹法检测表达产物。结果：经异丙基硫代-β—D-半乳糖苷诱导表达，表达产物的蛋白量约为菌体总蛋白的27．08％，相对分子质量为56000左右。能与抗鼠14-3-3γ多克隆抗体特异性结合。结论：表明成功构建了GST-14—3—3γ融合蛋白载体，在大肠杆菌中有高效表达，并具有良好的免疫反应性。

英文摘要：

Aim： To study the expression and purification of recombined rat 14-3-3γ protein in prokaryotic cell using genetic engineering technique, Methods： 14-3-3γ cDNA codon domain was amplified from rat myocardial tissue by RT- PCR method and recombined into pGEX-KG plasmid expressing glutathione S-transferase（GST） fusion protein. Results： GST-14-3-3γ fusion protein was induced by IPTG and further purified by GST agarose to obtain a fusion protein with molecular weight of 56 000, The expressed protein accounted for 27.08% in the total proteins. It was revealed that the expressed protein could react with a specific polyclonal antibody for the 14-3-3γ protein. Conclusion： GST-14-3-3γ fusion protein has been successfully constructed,which highly expresses in E. coli JM109 with good immunoreactivity.