郑州大学学报（理学版）Issue(4)：94-98,5.DOI:10.3969/j.issn.1671-6841.2015.04.018
GST-NAP融合蛋白可溶性表达及柱上切割GST标签
Expression of Soluble GST-NAP Fusion Protein and GST-tag-cleavage on Column
摘要
Abstract
To obtain plenty of HP-NAP ( H. pylori neutrophil-activating protein) for research, NAP gene was amplified from pMAL-C2X-NAP vector by PCR, and the recombinant plasmid pGEX-NAP was con-structed. After verified by enzyme digestion and gene sequencing analysis, pGEX-NAP was transformed into E. coli BL21(DE3)and the soluble fusion protein GST-NAP was obtained by inducing with IPTG in low temperature. Then GST-NAP was purified with Glutathione Sefinose Resin, and cleavaged GST-tag with Prescission protease on column. In conclusion, recombinant plasmid pGEX-NAP was successfully constructed and soluble GST-NAP fusion protein was efficiently expressed in E. coli. High purity HP-NAP can be obtained by Prescission protease cleavage on column. NAP protein was recognized by Rabbit anti-NAP mAb using Western Blot.关键词
GST-NAP/重组质粒/蛋白表达/GST标签Key words
GST-NAP/recombinant plasmid/protein expression/GST-tag分类
生物科学引用本文复制引用
黄夏冰,康巧珍,傅国,汲振余,刘鑫..GST-NAP融合蛋白可溶性表达及柱上切割GST标签[J].郑州大学学报（理学版）,2015,(4):94-98,5.基金项目
重大新药创制科技重大专项基金项目,编号2012ZX09103301-022 （）
国家自然科学基金资助项目,编号U1204817,81373119 （）
