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
