摘要
目的对人EIF2B4基因新的可变剪接产物进行鉴定。方法应用长链RT-PCR技术扩增人外周血EIF2B4基因编码区全长序列并进行T克隆及序列测定,用常规RT-PCR对新型可变剪接产物进行验证。结果在EIF2B4基因转录产物中检测出正常变异体2和3(两者只相差3个碱基),同时发现一个外显子7缺失26bp的转录产物。结论利用长链RT-PCR结合T克隆-测序的方法,发现人EIF2B4基因的一个新型可变剪接产物。
Objective To characterize a alternatively spliced product from human EIF2B4 gene.Methods The entire coding sequence of EIF2B4 gene was amplified from total RNA of peripheral blood cells by long distance RT-PCR,products of which were inserted into pGEM-T cloning vector,and multiple alternatively spliced products of the gene were analyzed by sequencing.Results In EIF2B4 gene,both transcript variant 2 and 3 were detected,which were different by 3 base pairs.A novel spliced product with a deletion of 26 bp in exon 7 was also found.This product was further confirmed by conventional RT-PCR.Conclusion A novel spliced product from human EIF2B4 gene was found using long distance RT-PCR,T cloning and sequencing.
出处
《福建医科大学学报》
2012年第1期36-38,共3页
Journal of Fujian Medical University
基金
福建省重点科技项目(2010Y0045)
关键词
剪接体
内含子
真核细胞起始因子2B
基因
逆转录聚合酶链反应
分子
克隆
序列分析
spliceosomes
introns
eukaryotic initiation factor-2B
genes
reverse transcriptase polymerase chain reaction
cloning
molecular
sequence analysis
