人TRAF2新剪切体的鉴定与表达分析  

Identification and expression analysis of a novel splice variant of human tumor necrosis factor receptor associated factor 2

作　　者：罗红雨[1] 张波[2] 严杰[2] 李玲[2] 尹抗抗[2] 韩梅[3] 

机构地区：[1]中南大学湘雅二医院肝病研究中心,湖南长沙410011 [2]湖南中医药大学中医学院,湖南长沙410208 [3]湖南师范大学生命科学院教育部蛋白质化学与发育生物学重点实验室,湖南长沙410081

出　　处：《细胞与分子免疫学杂志》2011年第11期1176-1179,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(31071150)

摘　　要：目的:对TRAF2的新剪切体进行鉴定与表达分析。方法:以人脑库为模板,利用PCR扩增,电泳确定新剪切体的存在。再提取不同细胞与组织的总RNA,逆转录得到cD-NA后作为模板对两种不同剪切体的表达进行比较分析。结果:利用P1与P2引物对从人脑库扩增出1条约1 500 bp的条带,而利用P3与P4引物对则得到2条明显电泳带,通过测序表明其中1条包含TRAF2的第6外显子,另1条缺失了TRAF2的第6外显子。通过对新剪切体TRAF2Δ6与全长剪切体TRAF2的表达量比较表明在T47D中全长剪切体表达占优势;而在Hep3B、GC-1与MCF7中TRATΔ6剪切体表达占优势;在HepG2、HBL100、A549与HeLa细胞中未发现两种剪切体的明显表达;在PANCI与SW480中两种剪切体表达量相近。在胎大脑与一级神经胶质瘤中TRAF2Δ6表达占优,而二级与三级神经胶质瘤中则全长剪切体TRAF2表达占优势。结论:人体TRAF2基因除可表达全长TRAF2 mRNA外,还可通过可变剪切表达缺失第6外显子的TRAF2Δ6 mRNA,而且这两种剪切体的表达存在细胞与组织特异性。AIM： Identification of a novel splice variant of tumor necrosis factor receptor associated factor 2 and its expression analysis. METHODS. By PCR analysis, we identified the existence of a new splice variant of TRAF2 using human brain cDNA library as template. RNA isolated from various cell lines and tissues was subjected to RT-PCR and differential expression analysis of TRAF2 splice variants was carried out. RESULTS： An amplification of about 1 500 bp was found using P1 and P2 as the primers, we performed PCR using exon 6 flanking primers P3 and P4, yielding a full-length transcript of 268 bp （including exon 6） and a novel splice variant of 193 bp lacking the 75 bp of exon 6. The full-length transcript of TRAF2 was found to be dominant in T47D, glioma of grade Ⅱ and grade Ⅲ. However the novel splice lacking exon 6 was dominant in Hep3B, GC-1, MCF7, fetus brain and glioma of grade Ⅰ, the two splices of TRAF2 were similar in PANCI, Hek293 and SW480. We couldn＇ t find any expression of TRAF2 in HepG2, HBL100, A549 and HeLa. CONCLUSION： TRAF2 has a novel splice lacking 6 exon in human beside the full-length splice. The two splices are differential expression in tissues and cell lines.

关 键 词：TRAF2 剪切体 PCR 

分 类 号：R392.12[医药卫生—免疫学]