以Bacmid-杆状病毒-昆虫细胞系统表达人FGF-9

Expression of human fibroblast growth factor 9 in Bacmid baculovirus expression system

目的：在Ｂａｃｍｉｄ杆状病毒昆虫细胞系统中表达人ＦＧＦ９ 。方法：采用ＲＴＰＣＲ技术，自新鲜人脑胶质瘤组织获取人ＦＧＦ９ 全编码区ｃＤＮＡ，将其克隆入ｐＣＲＴＭ Ⅱ质粒及ｐＹＥＸ４Ｔ１ 真核表达质粒，经ＤＮＡ自动测序仪进行ＤＮＡ序列测定。将人ＦＧＦ９ ｃＤＮＡ 定向克隆入ｐＦａｓｔＢａｃ 质粒，进一步将其转座入Ｂａｃｍｉｄ 中，在昆虫细胞Ｓｆ９ 中进行表达，采用ＳＤＳＰＡＧＥ对表达产物进行分析。结果：在昆虫细胞表达系统中表达出人ＦＧＦ９ 重组蛋白。结论：人ＦＧＦ９ 在Ｂａｃｍｉｄ杆状病毒昆虫细胞系统中得到了表达。

Objective: To express human fibroblast factor 9(hFGF 9). Methods: The full length cDNA of hFGF 9 was isolated from human glioma by reverse transcription PCR. The specific cDNA was cloned into pYEX4T 1 plasmid. The insert was confirmed by DNA sequencing , subcloned into pFastBac donor vector and transposed into Bacmid in DH10Bac. The recombinant Bacmid was used to transform Sf9 cells. The supernatant and cellular lysate were analyzed by using SDS PAGE. Results: A specific protein in the cellular lysate of Sf9 containing hFGF 9 with MW 30 kD was found. Conclusion: hFGF 9 has been expressed in the Bacmid baculovirus expression system.