从Hep-2细胞克隆hFGF-2 cDNA及序列分析  

Molecular cloning and sequencing of human FGF-2 in Hep-2 cell

作　　者：王帮河[1] 何金生[2] 汪春兰[1] 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》2005年第2期138-140,共3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: 03023056)

摘　　要：目的　从Hep 2细胞(人喉鳞癌细胞)克隆人的FGF 2cDNA,用于下一步构建携带hFGF 2cDNA的非复制型腺病毒载体。方法　设计和合成引物,提取Hep 2细胞的总mR NA,通过RT PCR扩增出目的cDNA,然后将其重组到pGEM TEasy载体中送测序分析。结果　经基因测序分析表明,从Hep 2细胞克隆的hFGF 2基因序列与已报告的序列,除在ATG后的第24位碱基发生无意突变外(由G→A),其他序列则完全一致。结论　从Hep 2细胞的总mRNA中成功克隆出hFGF 2的基因片段。Objective To obtain the cDNA encoding of FGF-2 from Hep-2 cells for further research. Methods The primers were designed based on the human FGF-2 mRNA sequence. Total mRNA was isolated from Hep-2 cells, and RT-PCR was performed. The fragment was ligated into pGEM-T Easy vector and sequenced. Results The sequence of hFGF-2 cDNA obtained from Hep-2 cells by RT-PCR was homologous to the sequence in GenBank(accession: J04513). Conclusion From cultured Hep-2 cells hFGF-2 cDNA has been cloned successfully.

关 键 词：成纤维细胞生长因子2 克隆 分子 序列分析 DNA DNA 互补 

分 类 号：R977.6[医药卫生—药品] R394.3[医药卫生—药学]