一个新的NPCEDRG基因mRNA剪接变异体的克隆及鉴定

Cloning and identification of a new alternative splicing variant of NPCEDRG

目的:克隆并鉴定NPCEDRG基因mRNA剪接变异体。方法:利用5'-RACE、3'-RACE在CNE2细胞中对NPCEDRG基因mRNA剪接变异体进行扩增,T/A克隆入pMD20载体,所获得的阳性重组子经测序证实。结果:成功克隆得到一种新的NPCEDRG基因的mRNA剪接变异体V2,其TSS位于-23 nt处,其CDs为297 bp,编码一种由98个氨基酸组成的蛋白质,其分子量为10.4 kD,等电点为7.1。结论:利用5'-RACE、3'-RACE等技术成功克隆得到一种新的NPCEDRG基因的mRNA剪接变异体,为阐述NPCEDRG基因在鼻咽癌发生发展中的可能作用提供线索。

Objective To further characterize NPCEDRG transcripts,we cloned and identified alternative splicing variants of NPCEDRG in an NPC cell lines CNE2.Methods 5'-RACE,3'-RACE were used to amplify full-length cDNA sequence of mRNA variants of NPCEDRG from CNE2.The PCR product was then inserted into pMD20 vector.And the positive recombinants were finally confirmed by sequence analysis.Results A new AS variant of NPCEDRG was cloned and identified,which contains a 297 nucleotides CDs and encode a protein consist of 98 aa(MW 10.4,PI 7.1).Conclusion This study clarified the AS variants of NPCEDRG,and a new AS variant of NPCEDRG was successfully cloned and identified,which provide important clues to better understanding of the molecular mechanism of human NPCEDRG gene expression in nasopharyngeal epithelial cells.