摘要
目的:研究人酸性成纤维细胞生长因子(haFGF)cDNA在不同表达系统中的表达, 旨在得到能高效表达haFGF的表达系统。方法:采用PCR方法扩增人酸性成纤维细胞生长因 子(haFGF)cDNA,将该cDNA片段分别插入表达载体pKK223-3及pET3C的表达框架中,筛选 得到了重组质粒pKK223-3-haFGF及pET3C-haFGF,分别转化大肠杆菌JM109及BL21(DE3), 构建了表达菌株JM109 KK223-3-haFGF及BL21(DE3)/pET3C-haFGF,用IPTG诱导表达。结 果:SDS-PAGE的结果表明,表达菌株BL21(DE3)/pET3C-haFGF能高效表达人酸性成纤维细 胞生长因子,而JM109/pKK223-3-haFGF诱导后目标蛋白带不明显。结论:BL21(DE3)/pET3C 表达系统更适于表达人酸性成纤维细胞生长因子。
Objective: The expression of human acidic fibroblast growth factor (haFGF) cDNA in pKK and pET expression system was studied. Methods: HaFGF cDNA produced by PCR was separately cloned into the expression vector pKK223-3 and pET3C. The expression of haFGF in JM109/pKK223-3-haFGF and BL21 (DE3) /pET3C-haFGF was induced by IPTG. Results: The expression level of haFGF in BL21 (DE3) /pET3C-haFGF was much higher than that in JM109/pKK223-3-haFGF. Conclusion: The expression system BL21 (DE3) /pET3C is suitable for haFGF expression.
出处
《中山大学学报论丛》
2001年第3期26-30,共5页
Supplement to the Journal of Sun Yatsen University