摘要
目的构建人血管内皮生长因子受体(VEGFR)-3胞外1-3区基因原核表达体系。方法提取人胎盘组织上总RNA,经反转录制备cDNA;从Genebank获取该基因序列,设计引物进行聚合酶链反应(PCR)以扩增VEGFR-3胞外1-3区基因片段,回收和纯化PCR产物并将其插入pMD18T载体中进行克隆;测序正确后,将目的基因克隆入原核表达载体pBV220,对获得的原核表达质粒进行初步鉴定。结果构建了pBV220-VEGFR-3质粒表达载体,经DNA测序鉴定序列正确。结论应用基因工程技术,成功构建了VEGFR-3原核表达载体。
Objective Construct the prokaryotic expression Vector for the gene of human vascular endothelial growth factor reeeptor（VEGFR）-3 extracellular domain1-3. Methods We extracted total RNA from human placental tissue,then reverse transcribed into eDNA.According to the eDNA seque,ce of VEGFR-3 acquired from genbank, specific primers for PCR were designed.The purified PCR product was connected with T vector and sequenced.We subcloned whis PCR product into pBV-220 plasmid and identified it. Results The pBV220-VEGFR-3 expression plasmid was constructed successfully.Sequence identified was correct. Conclusion Applying tecnique of genetic engineering,the pBV220-VEGFR3 expression plasmid was constructed successfully.
出处
《中国药物与临床》
CAS
2006年第3期169-171,共3页
Chinese Remedies & Clinics
基金
国家自然科学基金资助项目
关键词
受体
血管内皮生长因子
基因
遗传载体
Receptors,vascular endothelial growth factor
Gene
Genetic vectors
