中国免疫学杂志2018，Vol.34Issue(5)：708-711,717,5.DOI:10.3969/j.issn.1000-484X.2018.05.013
His标记STAT4(565-748氨基酸)肽段融合蛋白原核表达及纯化
Prokaryotic expression and purification of His-tagged-STAT4 (565-748 amino acid) fusion protein
摘要
Abstract
Objective:To construct His-tagged peptide of human STAT4 (565-748 amino acid) expression vector and induce its expression in Escherichia coli,followed by purification.Methods:STAT4 gene fragment encoding C-terminal peptide of 565-748 amino acid was amplified by PCR using pEGFP-STAT4 as the template.The PCR product was inserted into prokaryotic expression vector pET-28a and was transformed into component E.coli BL21 cells.By isopropyl-β-D-thiogalactoside(IPTG) induction,fusion protein was found to be expressed in the inclusion body and was denatured by using the urea denaturation buffer followed by renaturation and purifi-cation.Finally the purified protein was confirmed by Western blot.Results:The STAT4 truncated gene encoding 565-748 amino acids peptide was amplified by PCR and inserted into pET-28a vector.After the recombined plasmid was transformed into component BL21, the His-tagged-STAT4 (565-748 amino acids) fusion protein was induced and obtained after denaturation,refolding,purification and dialysis.Conclusion:The eukaryotic expression vector containing the truncated human STAT4 gene encoding 565-748aa peptide has been successfully constructed and the fusion protein was obtained.关键词
STAT4/基因克隆/蛋白表达/融合蛋白纯化Key words
STAT4/Gene cloning/Protein expression/Fusion protein purification分类
医药卫生引用本文复制引用
贺美,徐艳,姚芳玲,邹纯,于颍,黎明..His标记STAT4(565-748氨基酸)肽段融合蛋白原核表达及纯化[J].中国免疫学杂志,2018,34(5):708-711,717,5.基金项目
湖南省自然科学基金重点项目(12JJ2016)和中南大学研究生科研创新项目(1053320171363)资助. （12JJ2016）
