可溶性成纤维细胞生长因子受体-1基因的克隆和在快速翻译系统中高效表达对成纤维细胞增殖抑制作用

Cloning of sFGFR1 and its high efficient expression in RTS and inhibition of fibroblast proliferation

目的 克隆可溶性成纤维细胞生长因子受体 1(sFGFR 1)基因 ,并在快速翻译系统(RTS)中高效表达相应蛋白。方法 培养SwissRat 3T3成纤维细胞株 ,提取总RNA ,用逆转录 聚合酶链反应 (RT PCR)方法获取鼠sFGFR 1cDNA片段 ,酶切后克隆到 pIVEX2 .3d载体并进行序列分析 ;采用RocheRTSProteinMaster 5 0 0系统 ,高效表达sFGFR 1蛋白并用Westernblot鉴定表达的蛋白。体外转染大鼠血管内皮细胞。收集培养液上清 ,检测sFGFR 1及成纤维细胞生长因子 2 (FGF 2 )对成纤维细胞增殖抑制作用。结果 克隆了sFGFR 1基因 ,测序证实序列正确 ;Westernblot证实sFGFR 1蛋白在RTS系统中高效表达。FGF 2对STS成纤维细胞增殖具有明显抑制作用。结论 克隆sFGFR 1基因在RTS系统获得高效表达。对 3T3成纤维细胞具有明显增殖抑制作用。

Objective To clone sFGFR-1 gene and express corresponding protein in RTS.Methods Swiss Rat 3T3 fibroblasts were cultured.Total RNAs were extracted and sFGFR-1 cDNA was obtained by RT-PCR.By using NcoI and SmaI,the fragments were cut and the products were cloned into pIVEX2.3d vector and sequenced.sFGFR-1 protein was expressed in RTS ProteinMaster 500 and the products were analyzed by Western blot.Results sFGFR-1 gene was cloned.Sequencing analysis confirmed the cloned sequence was right.Western blot revealed that sFGFR-1 was expressed highly efficiently.Conclusion sFGFR1 gene was cloned and expressed highly efficiently in RTS.