| 血管内皮细胞生长因子受体Flt—1胞外区基因的克隆及其在中国仓？… |
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| 引用本文： | 曾革非 张智清. 血管内皮细胞生长因子受体Flt—1胞外区基因的克隆及其在中国仓？…[J]. 病毒学报, 2000, 16(2): 127-130 |
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| 作者姓名： | 曾革非 张智清 |
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| 作者单位： | [1]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验 [2]中国预防医学科学院病毒学研究所病毒基因工程国家 |
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| 摘 要： | 朋原代培养的人脐静脉血管内皮细胞（ＨＵＶＥＣ）提取细胞总ＲＮＡ，采用逆转录ＰＣＲ（ＲＴ－ＰＣＲ）方法得到ＶＥＧＦ受体Ｆｌｔ－１胞外区前３个ＩｇＧ样区域ｃＤＮＡ片段（Ｆｌｔ－１ｎ３）。将获得的受体基因克隆到真核表达载体ｐｃＤ－ＮＡ３．１中，得到重组质粒ｐｃＤＮＡ３．１／Ｆｌｔ－１ｎ３，通过南体转染方法将其转入中国仓鼠卵巢细胞（ＣＨＯ），用Ｇ４１８筛选得到稳定表达目的蛋白的细胞砍隆。经固相结合实验筛选
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| 关 键 词： | VEGF Flt－1 胞外区 基因克隆 基因表达 |
Cloning of Extracellular Domain of Vascular EndothelialGrowth Factor Receptor Flt-1 and Its Functional Expression in CHO Cells |
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| ZENG Ge fei,ZHANG Zhi qing,ZHOU Xiao ming,ZHANG Li gou,CHE Tuan jie,AN Nai li,CHEN Ai jun,YAO Li hong. Cloning of Extracellular Domain of Vascular EndothelialGrowth Factor Receptor Flt-1 and Its Functional Expression in CHO Cells[J]. Chinese journal of virology, 2000, 16(2): 127-130 |
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| Authors: | ZENG Ge fei ZHANG Zhi qing ZHOU Xiao ming ZHANG Li gou CHE Tuan jie AN Nai li CHEN Ai jun YAO Li hong |
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| Abstract: | The N terminal first three IgG like domain of VEGF receptor Flt 1 (flt l n3 ) was cloned from primary culture of human umbilical vascular endothelial cells (HUVEC) by RT PCR method. Flt l n3 was functionally expressed in stably transfected CHO cells by using recombinant plasmid PCF3, which was constructed by inserting Flt l n3 cDNA into eukaryotic expression vector pcDNA3.1. The product secreted in the medium can specifically bind to rh VEGF 165 and inhibit rh VEGF 165 stimulated HUVEC proliferation. Our results also showed the expressed Flt l n3 can inhibit angiogenesis of chick embryo chorio allantoic membrane(CAM). |
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| Keywords: | vascular endothelial growth factor receptor Chinese hamster ovary cell functional expression |
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