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牙鲆RAG2 基因的质粒构建和体外原核表达
王先磊^1,2, 谭训刚¹, 张培军¹, 徐永立¹
1.中国科学院海洋研究所, 实验海洋生物学重点实验室;2.青岛国家海洋科学研究中心

摘要:

将牙鲆(Paralichthys olivaceus)RAG2 cDNA 序列插入到体外表达质粒pProEXTM HT, 在大肠杆菌(Escherichia coli) BL-21 中进行体外表达, 分析了诱导时间和IPTG 浓度对重组蛋白产生量的影响,确定了最佳诱导条件为: 诱导时间为3 h, IPTG 诱导浓度0.2 mmol/L。本研究同时对RAG2 重组蛋白的可溶性进行确认, 并利用BD TALONTM 金属亲和柱纯化了RAG2 蛋白。本研究结果为进一步深入研究RAG2 蛋白的生物学功能奠定了良好的基础。

关键词: 牙鲆(Paralichthys olivaceus) RAG2 重组纯化

DOI：

分类号:

基金项目:国家863 计划资助项目(2012AA092203); 国家自然科学基金项目(30800838); 山东省自然科学基金项目(Y2008E12)

Construction and expression of a prokaryotic vector of recombinant olive flounder RAG2

Abstract:

The full length of olive flounder RAG2 cDNA was ligated to plasmid pPROEXTM HT, and was transferred into Escherichia coli (BL-21) for recombinant protein expression. It showed that the protein expression was induced by IPTG. The optimal concentration of IPTG and optimal induction time were 0.2 mmoL/L and 3 h, respectively. The recombinant RAG2 protein was purified by BD TALONTM Metal Affinity Resins. This study will be useful for further research on the function of flounder RAG2.

Key words: Olive flounder (Paralichthys olivaceus) RAG2 recombination purification

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