CAJ | 학술논문

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1106 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。
목적：학정MYCT1기인적전록기시점병극륭해기인신적전록본，탐토해기인적결구화공능。방법이용이지MYCT1적보수서렬，응용5′-RACE기술학정전록기시점위치，결합3′-RACE병접득도신전록본적정학전장；연후이용생물신식학연건대신전록본여MYCT1적cDNA서렬급안기산서렬진행대비분석；최후이용RT-PCR적방법분석신전록본적세포표체보。결과성공극륭득도장1106 bp적신전록본MYCT1-TV，기전록기시점위우ATG상유140 bp처。MYCT1-TV여MYCT1재결구상무명현차이，병엄범표체우각개세포계。결론 MYCT1전록기시점적학정화신전록본MYCT1-TV적극륭，위진일보연구MYCT1기인적전록조공궤제급기인공능전정료실험기출。
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. Re-sults 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 experi-mental basis for the further exploration and understanding of the function and the transcriptional regulation mechanism of MYCT1.