基因重组牛碱性成纤维细胞生长因子的纯化和鉴定

Purification and Idntification of Recombinant Bovine basic Fibroblst Frowth Factor

牛碱性成纤维细胞生长因子（ｂＦＧＲ）ｃＤＮＡ在Ｅ．ｃｏｌｉ中表达，高压均质破碎菌体后，上清液以Ｈｅｐａｒｉｎ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ亲和层析、Ｃ１８反相ＨＰＬＣ分高纯化得到一分子量为２．２５×１０￣４的蛋白质，其等电点为９．３４，并能与抗牛ｂＦＧＦＭｃＡｂ反应；其氨基酸组成与ｂＦＧＦ文献值一致；活性分析结果表明此蛋白质不仅能促进ＢＡＬＢ／ｃ３Ｔ３细胞增值，ＥＤ＿（５０）＝０．８５ｎｇ／ｍｌ，而且能促进鸡胚尿囊膜微血管增生，所得蛋白为具有生物活性的重组碱性成纤维细胞生长因子。

Using DNA Recombination techniques,we expeessed bovine bFGF cDNA in E. coli.Purifiedwith Heparin-Sepharose CL-6B and Revense-Phase HPLC,a protein with a MW of2.25×10_4 was obtained from the extract of the expression bacterial culture,The pI of thisprotein is 9.34,reacted strongly with anti-bovine bFGF McAb,and it is closely similarcomposition to natural bFGF,Bioassay of activity showed that this protein not onlyhad mitogenic activity for cultured BALB/c 3T3 cell,(ED_(50)=0.85 ng/ml)but also induced newcapillary blood vessel growth in chorioallantoic membrane.It was concluded that whatobtained was recombinant bovine bFGF with the biological activity similar to natural bFGF.