梭鱼白细胞介素-22原核表达及活性蛋白的制备
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江苏省自然科学基金项目(BK2011418)、江苏省省属高校自然科学基金项目(10KJB240001)和盐城工学院人才引进项目(XKR2011007)


Prokaryotic expression and preparation of Liza haematocheila interleukin-22 protein with biological activity
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    摘要:

    根据梭鱼IL-22 cDNA 序列设计特异性引物,从梭鱼脾脏中扩增IL-22用于表达片段,连接至载体pUC57。分别以BamHⅠ和KpnⅠ对含有目的基因的质粒和pQE-30进行酶切,连接并转化到大肠杆菌M15中,以IPTG进行诱导表达。表达产物通过Ni-NTA亲和层析进行纯化。经SDS-PAGE对表达产物进行鉴定。SDS-PAGE显示该IL-22表达质粒经IPTG诱导后,IL-22重组蛋白主要在菌体培养上清中表达。通过优化蛋白表达及纯化方法,成功获得了梭鱼IL-22活性蛋白。

    Abstract:

    To construct a prokaryotic expression system of Liza haematocheila interleukin (IL)-22,the partial IL-22 gene was cloned from spleen using the specific primers by PCR and inserted into pUC57 vector.Then,the plasmid and pQE-30 vector were digested with BamHⅠ and KpnⅠ,ligated and sub-cloned into M15.The IL-22 protein was expressed by IPTG induction,purified by Ni-NTA and confirmed by SDS-PAGE.The SDS-PAGE result showed that after induced by IPTG,the IL-22 was expressed successfully in M15,and mainly expressed in supernatant.The results indicated that the IL-22 prokaryotic expression vector was constructed successfully and the recombinant protein possessed biological activity.

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王资生,齐志涛,黄贝,张启焕,仇明,邵荣.梭鱼白细胞介素-22原核表达及活性蛋白的制备[J].华中农业大学学报,2012,31(6):752-755

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  • 收稿日期:2012-04-05
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