广东海洋大学学报Issue(4)：43-48,6.
融合蛋白IL6-OmpW的质粒构建及表达纯化
Construction and Purification of Prokaryotic Expression of Fusion Protein IL6-OmpW
摘要
Abstract
The fusion gene IL6-(Gly4Ser)3-OmpW was constructed with the DNA fragments of the red snapper IL-6 gene and Vibrio harveyi outer membrane protein OmpW gene which from recombinant plasmid pMD18-T/IL6 and pMD18-T/OmpW by Gene-SOEing method. The full length product was then cloned into the prokaryotic expression vector pET-32a(+) for protein expression in Escherichia coli strain BL21(DE3). The molecular weight of expression fusion protein IL6-(Gly4Ser)3-OmpW was about 66.6 ku. The recombinant protein was high expressed under induction conditions of exposure at 37℃, in 0.2 mmol·L-1 of IPTG for 5 h. The fusion protein was purified using HisTrap™HP affinity column and the best elution concentration of imidazole was 400 mmol·L-1. The concentration of purified fusion protein was 480 µg·mL-1. Western-blot analysis showed that the recombinant fusion protein could be combined with mouse anti-His-Tag Mab, indicating that the aim protein was expressed successfully. These results could provide a foundation for further study of its biological activity.关键词
哈维氏弧菌OmpW/红笛鲷IL-6/基因融合/原核表达Key words
Vibrio harveyi OmpW/Lutjanus sanguineus IL-6/Gene-SOEing/Prokaryotic expression分类
生物科学引用本文复制引用
黄浦江,黄郁葱,简纪常,吴灶和,鲁义善,黄瑜,樊云霞..融合蛋白IL6-OmpW的质粒构建及表达纯化[J].广东海洋大学学报,2013,(4):43-48,6.基金项目
国家科技支撑项目（2012BAD17B02）；国家自然科学基金项目（41240041）；广东省科技厅国际合作项目 （）
