从Hep-2细胞克隆hVEGF165/VEGF121基因及序列分析  

Molecular cloning and sequencing of human VEGF165/VEGF121 in Hep-2

作　　者：王帮河[1] 汪春兰[1] 何金生[2] 赵宇[1] 汪红俊[2] 曹东升[1] 

机构地区：[1]安徽医科大学第一附属医院整形外科,合肥230022 [2]安徽医科大学免疫学教研室,合肥230032

出　　处：《安徽医科大学学报》2004年第3期182-185,共4页Acta Universitatis Medicinalis Anhui

基　　金：安徽省科技厅自然科学基金资助项目 (编号 :0 3 0 2 3 0 5 6)

摘　　要：目的　从Hep 2 (喉鳞癌细胞 )克隆hVEGF16 5、hVEGF12 1cDNA并重组到克隆载体中 ,用于下一步构建表达VEGF 16 5、VEGF 12 1的腺病毒载体。方法　设计和合成引物 ,提取Hep 2细胞的总mRNA ,进行逆转录 -聚合酶链反应 (RT PCR) ,扩增出两个目的cDNA ,并将其重组到pGEM TEasy载体中送测序。 结果　从Hep 2细胞的总mRNA中反转录扩增得到 5 88bp和 4 5 6bp的片段 ,经重组送测序证实两基因分别与GenBank中的 [GI:1990 90 6 4 ]及[GI :6 6 310 2 8]提供序列完全一致。结论　从Hep 2细胞我们成功获得VEGF16 5、VEGF12 1的cDNA并构建其克隆载体可供进一步研究。Objective To obtain the cDNA sequence encoding VEGF165/VEGF121 from Hep-2 cell for further research.Methods The primers were designed were based on the human VEGF mRNA sequence.Total mRNA was isolated from Hep-2 cells, and RT-PCR was performed.The two fragments were recombined into pGEM-T Easy vector, and sequenced by automatic sequence analyzer.Results The sequences of VEGF165 cDNA and VEGF121cDNA from Hep-2 by RT-PCR were completely identical to the sequences GenBank provides.Conclusion From cultured Hep-2 cells the two cDNAs have been cloned successfully.

关 键 词：喉肿瘤 癌 鳞状细胞 内皮生长因子 克隆 分子 序列分析 DNA VEGF HEP-2 

分 类 号：R739.65[医药卫生—肿瘤]