MYCT1新转录本的克隆与表达分析  

Cloning and Expression Analysis of a Novel MYCT1 Transcript

作　　者：符爽 孙开来[1] 富伟能[1] 

机构地区：[1]中国医科大学基础医学院遗传学教研室,沈阳110001 [2]附属盛京医院血液研究室,沈阳110022

出　　处：《中国医科大学学报》2014年第5期388-392,共5页Journal of China Medical University

基　　金：国家自然科学基金(81372876);国家自然科学基金青年基金(81300420)

摘　　要：目的确定MYCT1基因的转录起始点并克隆该基因新的转录本,探讨该基因的结构和功能。方法利用已知MYCT1的保守序列,应用5′-RACE技术确定转录起始点位置,结合3′-RACE拼接得到新转录本的精确全长;然后利用生物信息学软件对新转录本与MYCT1的cDNA序列及氨基酸序列进行对比分析;最后利用RT-PCR的方法分析新转录本的细胞表达谱。结果成功克隆得到长1 106 bp的新转录本MYCT1-TV,其转录起始点位于ATG上游140 bp处。MYCT1-TV与MYCT1在结构上无明显差异,并广泛表达于各个细胞系。结论 MYCT1转录起始点的确定和新转录本MYCT1-TV的克隆,为进一步研究MYCT1基因的转录调控机制及基因功能奠定了实验基础。Objective To identify the transcriptional start site and clone a novel transcript variant of MYCT1（Myc target 1）for further study of its structure and function. Methods Transcriptional start site was confirmed using MYCT1 conserved sequence by 5′- RACE method and a novel MYCT1 isoform was cloned by splicing with 3′-RACE PCR product. Then,the cDNA or amino acid sequence between MYCT1 and its isoform was compared using bioinformatics server. Finally,the expression profile of this novel transcript in different cell lines was detected through RT-PCR. Results A 1 106 bp transcript named MYCT1-TV（Myc target 1 transcript variant 1）was successfully cloned,and its transcriptional start site was confirmed which located at 140 bp upstream of the ATG start codon of MYCT1-TV. MYCT1-TV shows no obviously structural difference with MYCT1 and is widely expressed in various cell lines. Conclusion The transcriptional start site analysis and MYCT1-TV cloning provide an experimental basis for the further exploration and understanding of the function and the transcriptional regulation mechanism of MYCT1.

关 键 词：MYCT1 MYCT1-TV 喉癌 RACE 

分 类 号：R394.3[医药卫生—医学遗传学]