斑马鱼SFPQ蛋白的原核表达及纯化

Prokaryotic Expression and Purification of Zebrafish SFPQ

Splicing Factor Proline and Glutamine Rich简称为SFPQ,是广泛表达在脊椎动物中的一类蛋白。它作为抑癌基因,对生物体内肿瘤发生具有重要的调控作用。目前对于SFPQ的研究主要集中在人和小鼠上,对于斑马鱼SFPQ的研究还比较少。为了获得原核表达的SFPQ蛋白进行下一步研究,构建了斑马鱼SFPQ-p ET28a重组质粒,然后转化到BL21化学感受态细胞中,探索用IPTG进行低温诱导蛋白的最适条件,然后用Ni-NTA琼脂糖镍柱纯化SFPQ蛋白。实验结果表明,当IPTG浓度为0.75 mmol/L,诱导时间为10 h时,蛋白表达量最大,Western blot结果表明纯化得到的斑马鱼SFPQ蛋白可以和抗体特异性结合,说明纯化得到的斑马鱼SFPQ蛋白可以用于下一步实验。

Splicing Factor Proline and Glutamine Rich（SFPQ）is a protein that expresses widely in vertebrates. As a tumor suppressor gene, it plays the vital regulation role in tumorigenesis of living organisms. So far the researches of SFPQ were focused on human being and mice, little has been known about the SFPQ of zebrafish. In order to acquire the prokaryotic-expressed SFPQ protein for further research, recombinant p ET28a-SFPQ plasmid of zebrafish was constructed and transformed into the chemical competent cells of BL21. The optimal condition was explored while IPTG was applied to induce the expression of zebrafish SFPQ protein in low-temperature, and the SFPQ protein was purified by Ni-NTA agarose nickel column. The results demonstrated that the expression of the protein was the most while inducing for 10 h by 0.75 mmol/L IPTG at 27℃. The results of Western blot revealed that the purified SFPQ protein specifically bound with the antibody, indicating that the purified zebrafish SFPQ protein may be utilized in the further experiments.