【摘 要】
：
从人胚肺二倍体细胞KMB17中抽提总RNA,经RT-PCR扩增获得编码人角质细胞生长因子-2(keratinocyte growth factor-2,KGF-2)的cDNA.克隆于硫氧环蛋白表达载体pThioHisA,序列分析
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从人胚肺二倍体细胞KMB17中抽提总RNA,经RT-PCR扩增获得编码人角质细胞生长因子-2(keratinocyte growth factor-2,KGF-2)的cDNA.克隆于硫氧环蛋白表达载体pThioHisA,序列分析表明与文献报道一致.经IPTG诱导,在大肠杆菌BL21中实现高效表达,表达量可达菌体总蛋白10%～15%.菌体超声破碎,上清经CM-Sepharose FF阳离子交换,Heparin-Sepharose亲和层析,Superdex 75凝胶过滤层析纯化得到重组人KGF-2,纯度高于95%.生物活性分析表明,它能够促进成纤维细胞NIH/3T3的增殖,诱导鸡胚背根神经结神经轴突的生长,促进鸡胚尿囊膜血管生成.研究结果表明,获得了95%纯度的具有生物学活性的重组人KGF-2,为进一步的基础与应用研究提供了基础.
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