KDR是血管内皮生长因子(VEGF)的主要受体之一,它在介导VEGF刺激内皮细胞增殖及血管通透性等生物学活性中起重要作用.为了获得人重组的有VEGF结合活性的KDR,通过RT-PCR从人胎儿脐静脉内皮细胞(EC)扩增出编码KDR胞外Ⅰ~Ⅳ区片段,将其克隆在谷胱甘肽转移酶(GST)融合蛋白表达载体pGEX2T中,并在大肠杆菌中获得稳定表达.表达的融合蛋白GST-KDR以包涵体形式存在,其分子质量约66 ku左右.经碱变性法大量提取,及制备胶电泳获得纯化的KDR,为进一步的研究奠定了基础.
VEGF receptor, KDR, is one of the main VEGF receptors, and it plays important role in VEGF stimulating endothelial cell (EC) proliferation and vascular permeation. To obtain active recombinant KDR which can bind VEGF in vitro, RT-PCR was used to clone KDR (Ⅰ~Ⅳ) DNA fragment from human umbilical vein EC. The cloned KDR (Ⅰ~Ⅳ) fragment was identified with enzyme digestion and sequencing and then cloned into fusion protein expression vector pGEX2T. The GST-KDR fusion proteins were expressed in E.coli XL1-blue after inducing by IPTG. The fusion proteins were extracted from bacterial inclusion body with basic denature method and purified with preparing SDS-PAGE gel followed by electric-elution.
宋述梅,寿成超. VEGF受体KDR结合区的克隆与表达[J].生物化学与生物物理进展,1999,26(5):481-484
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