新的含有11q13.5HERV-Wgag序列的内含子转录子的鉴定  

Identification of a Novel Intronic Transcript Containing 11q13.5 HERV-W gag Sequence

作　　者：王杰[1] 姚源蓉[1] 

机构地区：[1]天津医科大学总医院神经内科,天津市神经病学研究所,300052

出　　处：《天津医药》2010年第1期1-3,共3页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:30800356);天津市自然科学基金资助项目(项目编号:07JCYBJC16200);教育部留学回国人员科研启动基金(项目编号:教外司留[2008]890号);天津医科大学总医院引进人才科研启动基金

摘　　要：目的:鉴定位于染色体11q13.5的含有HERV-Wgag序列的内含子转录子,探讨这一新的转录子对宿主基因PTD015选择性剪切的调控作用。方法:采用半巢式PCR和降落PCR扩增目的片段,克隆、测序、构建载体、转染JEG3细胞,采用实时PCR检测PTD015选择性剪切mRNA的水平。结果:鉴定了一个位于基因PTD015第二内含子上长1739bp的转录子,该转录子包含755bp11q13.5HERV-Wgag序列、527bp5′长末端重复序列和11q13.5HERV-W5′端457bp片段。此内含子反义转录子质粒的转染使JEG3细胞PTD015选择性剪切mRNA的水平明显降低。结论:源于基因PTD015第二内含子含有HERV-Wgag序列的反义转录子可调节宿主基因PTD015的选择性剪切。Objective：To identify the novel intronic transcripts containing 11q13.5 HERV-W gag sequence,and explore the modulation of the transcripts on the alternative splicing of host gene PTD015. Methods：Half-nested PCR and touchdown PCR were used to amplify the target transcripts. The transcripts were cloned and sequenced. The plasmids inserted with the target transcripts were transfected into JEG3 cells and the alternative spliced mRNA levels of PTD015 were measured with real time PCR. Results：A 1 739 bp novel intronic transcript containing 755 bp 11q13.5 HERV-W gag sequence,527 bp 5＇ long terminal repeat and 457 bp sequence on the 5＇-end of 11q13.5 HERV-W was identified. The intronic antisense transcripts significantly down-regulated the alternative spliced mRNA levels of PTD015. Conclusion：The intronic antisense transcripts originating from the second intron of gene PTD015 could modulate the alternative splicing of the host gene PTD015.

关 键 词：染色体 人 11对 内源性逆转录病毒 基因 内含子 RNA剪接 

分 类 号：R734.2[医药卫生—肿瘤] R733.7[医药卫生—临床医学]