CAJ | 학술논문

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cDNA，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。
목적：전록인자Pax-8재배태발육급종류발생중구유중요작용，본연구지재극륭급감정소서Pax-8기인아형，병검측기시공표체。방법：종야생형C57BL/6태서적심장중제취총RNA（Trizol법），반전록성cDNA，응용Q5초보진매진행목적기인확증，병극륭지진핵표체질립，진행매절감정급측서감정。결과：재소서발육불동시기，제이지전장Pax-8a외，발현3개신적소서Pax-8선택성전접체，분별명명위Pax-8b（제4외현자결실）、Pax-8c（제4화제11외현자결실）、Pax-8d（제4화제9외현자결실）。결론：소서심장중Pax-8존재사충선택성전접체，차재불동발육시기차이표체。
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.