宫颈癌组织中转录因子Skn-1 a cDNA的克隆及表达  

cDNA cloning and expressing of human Skn-1a cDNA from cervical carcinoma tissue

作　　者：王艳萍[1] 马正海[1] 余勐[1] 热西旦.尼格买提 马彩玲[2] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院分子生物学重点实验室新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046 [2]新疆医科大学第一附属医院妇产科,新疆乌鲁木齐830054

出　　处：《新疆医科大学学报》2006年第5期397-400,共4页Journal of Xinjiang Medical University

基　　金：教育部科学技术研究重点项目(02171);新疆维吾尔自治区自然科学基金项目(200221103)

摘　　要：目的:从宫颈癌组织中扩增与上皮细胞分化和人乳头瘤病毒(HPV)基因表达相关的转录因子Skn-1acDNA,进而在真核细胞中表达该转录因子以了解其功能。方法:从宫颈癌组织中提取总RNA,以此为模板,RT-PCR扩增Skn-1acDNA片段,将该cDNA片段克隆至pMD18-T载体并进行序列分析,构建Skn-1a真核表达载体,并转染Hela细胞,检测其表达情况。结果:RT-PCR扩增得到约1 300 bp的cDNA片段,将其克隆至pMD18-T载体,DNA测序表明,得到片段长度为1 308 bp,为转录因子Skn-1a基因编码的cDNA,获得的Skn-1acDNA序列与已公布的Skn-1acDNA序列具有99.85%同源性,有1个碱基的差异,但并未导致编码氨基酸的改变,继而构建了Skn-1a真核表达载体pcDNA3/Skn-1a,其转染Hela细胞后可正确表达。结论:转录因子Skn-1a在宫颈癌组织中有一定量的表达,并在体外培养的Hela细胞中获得表达。Objective： To clone cDNA of human transcription factor .Skn-1a （related to epidermal differentiation and HPV gene expression） gene from cervical carcinoma tissue and construct the eukaryotic expression vector of the Skn-1a cDNA. Methods： The Skn-1a cDNA fragment was amplified by RT-PCR from total RNA that was extracted from cervical carcinoma tissue,the PCR product was cloned into pMD18-T vector and the nucleotide sequence was analyzed, subsequently, the eukaryotic expression vector of the Skn-1a cDNA was constructed, and transfected Hela cells, the expression of Skn -1a was examined. Results： A cDNA fragment about 1300 bp was amplified by RT-PCR,the result of sequencing showed the fragment was 1 308 bp in length, containing a cDNA coding human Skn-1a . The nuclcotide sequence homology to the previously published sequence of human .Skn-1a was 99.85%, there was one conversion in nucleic acid sequences, but the conversion didn＇t induce the change of coding amino acid; the eukaryotic expression vector of Skn-1a can express correctly after transfecting Hela cells. Conclusions： The results suggested that Skn- 1a gene expresses at some level in the cervical carcinoma tissue, and Skn-1a cDNA can express in vitro culture Hela cells. The research provide basis for further exploring the relationship between Skn-1a and cervical carcinoma.

关 键 词：宫颈癌 Skn-1a 克隆 表达 

分 类 号：R737.33[医药卫生—肿瘤] R-34[医药卫生—临床医学]