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为了获得有活性的雷竹Pv SOC1蛋白(Phyllostachys violascens SUPPERSSOR OF CO OVEREXPRESSION 1),并进一步讨论其结构形态,通过原核表达方法得到Pv SOC1的可溶性重组蛋白,以p ET-GST作为表达载体,大肠杆菌E.coli Rosetta作为宿主菌株,超表达全长和IKC结构域的雷竹Pv SOC1蛋白。发现在37℃条件下,菌液浓度D(600)值达到0.4~0.6时进行诱导,控制IPTG终浓度为0.4 mmol/L,诱导目的蛋白表达5 h,可以获得可溶的IKC结构域GST融合蛋白。采用TEV(tobacco etch virus protease)酶切技术、配合GST亲和层析柱及分子层析色谱柱,去除标签蛋白并纯化目的蛋白。所获得的高纯度IKC结构域Pv SOC1蛋白经过对比分析发现其以八聚体形式存在。
In order to obtain the active Pv SOC1 protein (Phyllostachys violascens SUPPERSSOR OF CO OVEREXPRESSION 1) and to further discuss its structural morphology, we obtained the soluble recombinant protein of Pv SOC1 by using prokaryotic expression method. Using p ET-GST as the expression vector, E. coli Rosetta as a host strain overexpression of full-length and IKC domains of Thompson pv SOC1 protein. The results showed that under the condition of 37 ℃, the concentration of D (600) reached 0.4-0.6, the final concentration of IPTG was 0.4 mmol / L and the expression of target protein was induced for 5 h, and the soluble IKC domain GST fusion was obtained protein. Using TEV (tobacco etch virus protease) digestion technology, combined with GST affinity chromatography and molecular chromatography column, remove the tagged protein and purify the target protein. The obtained high purity IKC domain Pv SOC1 protein was found to be in octamer form by comparative analysis.