Cloning and Expression Analysis of a Novel MYCT1 Transcript / 中国医科大学学报

ABSTRACT

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.