草原短尾羊TBXT基因RACE克隆及胚胎各个时期表达量分析  

RACE Cloning of TBXT and Expression of Embryo in Different Stages of Prairie Short Tail Sheep Analysis

作　　者：杨光 霍雨佳 智达夫[1,2] 苏红 杨宏新 窦傲蕾 曹贵方 Yang Guang;Huo Yujia;Zhi dafu;Su Hong;Yang Hongxin;Dou Aolei;Cao Guifang(School of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot Inner Mongolia 010018,China;Key Laboratory of Basic Veterinary Medicine of Inner Mongolia Autonomous Region,Hohhot Inner Mongolia 010018,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]内蒙古自治区基础兽医学重点实验室,内蒙古呼和浩特010018

出　　处：《中国畜牧杂志》2022年第4期104-110,共7页Chinese Journal of Animal Science

基　　金：国家自然科学基金(3186130045);内蒙古自治区科技计划(2019GG241)。

摘　　要：为鉴定草原短尾羊TBXT mRNA在胚胎发育中的表达趋势,分析其TBXT cDNA全长序列。本研究使用实时荧光定量PCR(Quantitative Real time PCR)方法对草原短尾羊14、16、20 d和25 d胚胎中TBXT mRNA相对表达量进行分析,之后使用RACE(Rapid Amplification of cDNA Ends)技术克隆TBXT cDNA全长。结果显示:TBXT mRNA在16 d胚胎中相对表达量最高,草原短尾羊TBXT cDNA全长为2426 bp(GenBank登录号:MZ146381),其中开放阅读框(ORF)为1335 bp,编码444个氨基酸,3’非编码区(3’UTR)和ployA尾长度为763 bp,5’非编码区(5’UTR)长度为328 bp;系统进化树显示草原短尾羊Brachyury与山羊进化关系最近,与羊驼关系最远,氨基酸同源性比对结果显示不同物种之间Brachyury同源性很高,表明在脊索动物进化过程中Brachyury功能相对保守。This paper aimed to identify the expression trend of TBXT gene in prairie bobtail sheep during embryonic development and analyze the full-length cDNA sequence of prairie bobtail sheep TBXT.The relative expression of TBXT mRNA in 14 day,16 day,20 day,and 25 day embryos of prairie bobtail sheep was analyzed by qPCR method.Prairie bobtail sheep TBXT gene was cloned by 5’and 3’RACE from 16 day embryos and analyzed by the bioinformatics.The results indicated that TBXT mRNA expressed the highest level in 16 day embryos,The full length of TBXT cDNA of prairie bobtail sheep was 2426 bp(GenBank accession NO.MZ146381),an open reading frame is 1335 bp encoding 444 amino acids,3’noncoding region and polyA tail length is 763 bp,5’non-coding region length is 328 bp;phylogenetic tree shows prairie bobtail sheep Brachyury has the closest evolutionary relationship with goats,and the farthest relationship between alpaca.Amino acid homology comparison results show that Brachyury has high homology between different species.Brachyury functions are relatively conservative during the evolution of chordates.

关 键 词：草原短尾羊 实时荧光定量 RACE TBXT基因 

分 类 号：S826.2[农业科学—畜牧学]