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| 人血管内皮细胞生长因子cDNA克隆和在大肠杆菌的高效表达 | |
| 引用本文： | 何延政,刘建平,谢毅,张培华. 人血管内皮细胞生长因子cDNA克隆和在大肠杆菌的高效表达[J]. 中国普外基础与临床杂志, 2001, 8(2): 72-74 |
| 作者姓名： | 何延政 刘建平 谢毅 张培华 |
| 作者单位： | 1. 泸州医学院附属医院血管外科 四川泸州 646000 2. 复旦大学遗传工程国家重点实验室 上海 200433 3. 上海第二医科大学附属第九人民医院血管外科 上海 200011 |
| 基金项目： | 上海教育委员会基金（95GB04） |
| 摘 要： | 目的 在大肠杆菌高效表达人血管内皮生长因子蛋白质。方法 用PCR从人胎儿脑cDNA文库扩增血管内皮细胞生长因子（VEGF）cDNA，得到517bp的DNA片段。扩增片段重组到M13mP18中，经测序证实为VEGF165cDNA。将该片段重组到PRL621温控表达质粒中，在大肠杆菌表达20kd的重组蛋白。结果 该表达产物占菌体总蛋白的35%，其N-端15个氨基酸序列与天然VEGF165蛋白相应序列一致。结论 工程菌TG1/PRL621/VEGF高效表达人VEGF165蛋白质。 |
| 关 键 词： | 血管内皮细胞生长因子 克隆 表达 cDNA 大肠杆菌 |
| 文章编号： | 1007-9424(2001)02-0072-03 |
| 修稿时间： | 1999-12-25 |
CLONING cDNA OF HUMANVASCULAR ENDOTHELIAL GROWTH FACTOR AND HIGH LEVEL EXPRESSION IN ESCHERICHIA COLI |
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| HE Yan-zheng,LIU Jian-ping,XIE Yi,et al.. CLONING cDNA OF HUMANVASCULAR ENDOTHELIAL GROWTH FACTOR AND HIGH LEVEL EXPRESSION IN ESCHERICHIA COLI[J]. Chinese Journal of Bases and Clinics In General Surgery, 2001, 8(2): 72-74 | |
| Authors: | HE Yan-zheng LIU Jian-ping XIE Yi et al. |
| Affiliation: | HE Yan-zheng*,LIU Jian-ping,XIE Yi,et al.*Research Division of Vascular Surgery,Hospital of Luzhou Medical College,Luzhou 646000 |
| Abstract: | Objective To be expressed human vascular endothelial growth factor (VEGF) recombinant protein in Escherichia Coli in high level. Methods VEGF was amplified from human fetal brain cDNA library, the amplified fragment was inserted into M13mP18 and confirmed to be VEGF165cDNA by restriction mapping and DNA sequencing, then it was combined with an expression vector PRL621. This recombinant plasmid overexpressed a 20kd recombinant protein in E.Coli(TG1), the protein was isolated and purifed from E.Coli, and initially renatured. Results The overexpressed recombinant protein was 35% of the total cell protein, the sequence of its first 15-N terminal amino acid was identrical to that of the human natural VEGF protein, Chorioallantoic membrane(CAM) assay showed that the rhVEGF promated new capillary vessels formation. Conclusion The genetic engineering Escherichia Coli can express human vascular endothelial growth factor in high level. |
| Keywords: | Vascular endothelial growth factorCloning Expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |