人尿酸转运蛋白基因的克隆与序列分析  

Cloning and sequence analysis of human uric acid transporter gene

作　　者：洪权[1] 吴镝[1] 陈香美[1] 侯剀[1] 

机构地区：[1]解放军总医院肾病中心暨肾病重点实验室,北京100853

出　　处：《第一军医大学学报》2005年第6期623-625,629,共4页Journal of First Military Medical University

基　　金：国家自然科学基金"创新研究群体"项目(30121005);国家"973"项目(G2000057000);国家自然科学基金(30100062)~~

摘　　要：目的获得编码人尿酸转运蛋白(humanuricacidtransporter,hUAT)的全长基因。方法从人肾小管上皮细胞株(HK-2)中提取总RNA,根据Genbank中hUAT基因序列设计特异性引物,然后通过RT-PCR法扩增目的片段。所得目的片段酶切后与载体pEGFP-C1连接,经酶切及序列分析鉴定。结果成功扩增出980bp的hUAT全长基因,克隆到pEGFP-C1载体后酶切分析结果与预期一致,序列分析结果与Genbank上公布的hUAT序列完全一致。结论成功克隆出hUAT基因,序列分析结果完全正确,为对该基因进一步研究奠定了基础。Objective To obtain full-length human urate transporter (hUAT) gene. Methods Primers was designed according to the sequence of hUAT reported in Genbank. The target fragments were obtained by reverse transcriptional (RT) PCR from the mRNA extracted from human renal tubular epithelial cell lines (HK-2), followed by cloning into pEGFP-C1 plasmid. The cloned fragment was subjected to restriction mapping and sequence analysis for confirmation. Results and Conclusion hUAT gene with correct sequence is successfully cloned from HK-2 cells. The restriction maps and sequence analysis of the selected clones were consistent with the sequence reported in Genbank.

关 键 词：尿酸转运蛋白 逆转录-聚合酶链反应 克隆 序列分析 

分 类 号：R346[医药卫生—基础医学]