胸腺素α原及胸腺九肽在体内免疫方面有重要作用 .根据大肠杆菌遗传密码的偏爱性 ,设计并人工合成了胸腺素α原及胸腺九肽融合蛋白的特异引物 .以胸腺素α原cDNA为模板 ,应用PCR及分子克隆技术构建了该融合蛋白的编码基因 ,将其插入pBV2 2 0质粒 ,转化大肠杆菌 ,通过温度诱导使该融合蛋白高效表达 ,并对表达产物进行了高效特异性纯化 .运用MTT法初步测定了该融合蛋白的活性 .结果表明 ,该融合蛋白对氢化可的松诱导的胸腺细胞损伤有一定的保护作用 ,为今后进一步研究和应用该融合蛋白奠定了基础
Abstract
Prothymosin α and thymulin are important immune factors in vivo . The special primers were designed and synthesized according to the codon bias of E.coli. Fusion protein gene of the two proteins was amplified by PCR, inserted into prokaryotic expression vector pBV220 and transferred in E.coli . The over expression product induced by temperature shift was purified effectively and assayed for its biological activity by MTT. The data indicated that the fusion protein had protective effect on injury of thymocyte induced by hydrocortisol. The high expression of fusion protein may provide a material to explore its function and clinical usage in future.
关键词
胸腺素α原 /
胸腺九肽 /
融合蛋白 /
基因表达 /
纯化
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Key words
prothymosin α /
thymulin /
fusion protein /
gene expression /
purification
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参考文献
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