蜂毒肽
重组DNA
赫拉
融合蛋白
分子生物学
Myc标签
亲和层析
标志标签
生物活性
紫胶操纵子
蛋白质A/G
蛋白质纯化
化学
靶蛋白
体外
生物化学
生物
色谱法
肽
酶
基因
摘要
Objective To prepare purified recombinant melittin- 88ArgIL-2 chimeric protein and analyze its biological activity.Methods Transform the constructed recombinant plasmid pGEX-4T-2/melittin- 88ArgIL-2 to E.coli DH5α for expression under induction of IPTG.Purify the expressed product by ion exchange and affinity chromatography and identify by SDS-PAGE.Determine the inhibitory effect of expressed chimeric protein on the proliferation of Hela cells by MTT method.Results The purity and protein concentration of expressed soluble protein after purification reached 95% and 0.5 g/L respectively.The purified chimeric protein,at a concentration of 20 μg/L,showed significant inhibitory effect in vitro on the proliferation of Hela cells.Conclusion The purified recombinant melittin- 88ArgIL-2 chimeric protein was successfully prepared and showed biological activity.It laid a foundation of scale-up of pilot procedure and development of purification procedure of the chimeric protein.
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