三个新的小鼠Pax-8剪接体的克隆及时空表达  

Cloning of three newly-identified alternatively splicing isoforms of mouse Pax-8 and their spatial and temporal expression patterns

作　　者：宋丽娟[1] 倪云杰 林振浩 侯良磊 吴漪皓[1] 胡欢欢[1] 黄晓燕[2] 杨德业[1,2] 

机构地区：[1]温州医科大学附属第一医院心内科,浙江温州325015 [2]杭州师范大学附属医院心内科,浙江杭州310015

出　　处：《温州医学院学报》2015年第4期240-242,247,共4页Journal of Wenzhou Medical College

基　　金：国家自然科学基金资助项目(81270230)

摘　　要：目的:转录因子Pax-8在胚胎发育及肿瘤发生中具有重要作用,本研究旨在克隆及鉴定小鼠Pax-8基因亚型,并检测其时空表达。方法:从野生型C57BL/6胎鼠的心脏中提取总RNA(Trizol法),反转录成c DNA,应用Q5超保真酶进行目的基因扩增,并克隆至真核表达质粒,进行酶切鉴定及测序鉴定。结果:在小鼠发育不同时期,除已知全长Pax-8a外,发现3个新的小鼠Pax-8选择性剪接体,分别命名为Pax-8b(第4外显子缺失)、Pax-8c(第4和第11外显子缺失)、Pax-8d(第4和第9外显子缺失)。结论:小鼠心脏中Pax-8存在四种选择性剪接体,且在不同发育时期差异表达。Objective：Pax-8 gene plays a pivotal role in embryonic development and tumorgenesis. This study aims to identify and clone different alternatively splicing isoforms of mouse Pax-8, and to detect their spa-tial and temporal expression patterns. Methods：Hearts from wild-type prenatal and postnatal mice of C57BL/6 background were carefully excised and total RNA was extracted using Trizol reagent. The cDNA fragment of Pax-8 was reverse transcribed and ampliifed by Q5 Taq polymerase and cloned into pcDNA3.1（-） vector. The re-combinant vectors were validated by enzyme digestion and sequencing. Results：In comparison to their cognate full-length Pax-8a, three alternatively splicing isoforms of mouse Pax-8, designated Pax-8b （exon4 deleted）, Pax-8c （exon 4 and 11 deleted） and Pad-8d （exon 4 and 9 deleted）, were identiifed in different periods of embryogen-esis and successfully cloned. Conclusion：Mouse heart has four alternatively splicing isoforms of mouse Pax-8 that express in different spatial and temporal expression patterns. Such ifndings open a bright avenue for the further functional study.

关 键 词：Pax-8基因 剪接体 时空表达 克隆 小鼠 

分 类 号：Q785[生物学—分子生物学]