摘要
目的构建原核表达质粒pPROEXTMHTbGMCSF/IL15并在大肠杆菌DH5α中表达,以期获得具有GMCSF/IL15双重活性的融合蛋白。方法利用分子生物工程技术,构建重组原核表达载体pPROEXTMHTbGM15,并转化进大肠杆菌DH5αIPTG诱导表达4h后,经SDSPAGE、免疫印迹证实为GM15融合蛋白。结果重组载体经转化大肠杆菌DH5α后,诱导表达出相对分子质量约为33kD左右的融合蛋白。结论构建了pPROEXTMHTbGMCSF/IL15融合基因表达系统,并诱导表达出GM15融合蛋白。
Objective Construct pPROEXTM HTb-GM-CSF/IL-15 and express in DH5α to obtain the GM-15 fusion protein, which may have double activity. Methods Using molecular biological techniques, a recombinant vector pPROEXTMHTb-GM-CSF/IL-15 was constructed, and express fusion protein inducing by IPTG 4 h. The fusion protein was vetified by SDS-PAGE, Western blot. Results The fusion protein with relative molecular mass of 33 kD was yielded in DH5α by induced. If possessed double activity of anti GM-CSF and anti IL-15 by western blot assay. Conclusion The successful construction of the expression vector laid a foundation of largly product and clinical use of the fusion protein.
出处
《福建医科大学学报》
2006年第2期111-113,共3页
Journal of Fujian Medical University
基金
福建省卫生厅医学创新基金资助项目(2001CX12)
关键词
GM—CSF
白细胞介素15
重组融合蛋白质类
质粒
基因表达
granulocyte-macrophoge colong-stimulating factor
interleukin-15
recombinant fusion protein
plasmids
gene expression
