定量诱导表达PRDM1永久细胞系的建立

目的建立可定量诱导表达PRDM1的永久细胞系,为进一步研究PRDM1的功能创建平台。方法将野生型PRDM1基因的cDNA全长克隆到pMEP4表达质粒中,电穿孔法将pMEP4-PRDM1瞬时转染到细胞系SALT3,经潮霉素B筛选后挑选可增殖的单克隆细胞并大量扩增,在不同浓度的硫酸镉（CdSO4）诱导下检测PRDM1蛋白的表达状况。结果成功构建了pMEP4-PRDM1表达载体,在最佳电穿孔条件下将其转染到SALT3细胞,经潮霉素B筛选后扩增出6个细胞克隆。CdSO4诱导后运用免疫印迹法验证了PRDM1蛋白的表达与诱导剂CdSO4存在量效关系。结论运用pMEP4载体可建立定量诱导表达目的基因PRDM1的永久细胞系,为进一步研究PRDM1的功能奠定了基础。

Establishment of quantitatively inducible PRDMI-expressing stable cell line

Objective To establish the quantitatively inducible PRDM1-expressing stable cell line in order to create a platform for the study of PRDM1 function. Methods Full-length PRDM1 cDNA was inserted into the multiple cloning site of pMEP4 vector. A pMEP4-PRDM1 expression vector was instantaneously transfected into SALT3 cell line by electroporation, followed by hygromycin B selection. Hygromycin-resistant stable clones were expanded and PRDM1 protein expression was detected at different doses of CdSO4 by Western blot. Results The pMEP4-PRDM1 expression vector was successfully constructed and subsequently transfected into the SALT3 cell line under optimal electroporation conditions. Six hygromycin-resistant stable clones were established and expanded. The PRDM1 protein expression in these stable cell lines was proved to be quantitatively correlated with the dose of CdSO4. Conclusion Quantitatively inducible PRDM1-expressing stable cell line can be established using the pMEP4 vector, thus laying a good foundation for further analysis of the function of this gene.