奶山羊垂体同型框转录因子2基因(PITX2)分子克隆、序列分析及其mRNA表达规律  

Molecular Cloning, Sequence Analysis and m RNA Expression Profiling of Paired- liked Homodomain Transcription Factor 2 Gene(PITX2) in Dairy Goat(Capra hircus)

作　　者：张晓燕[1] 赵海谕[1] 张思欢 雷初朝[1] 陈宏[1] 蓝贤勇[1] 

机构地区：[1]西北农林科技大学动物科技学院/陕西省农业分子生物学重点实验室,杨凌712100

出　　处：《农业生物技术学报》2015年第7期905-916,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31172184);陕西省青年科技新星基金(No.2011kjxx64)

摘　　要：垂体同型框转录因子2(paired-liked homodomain transcription factor 2,PITX2)是Wnt/β-catenin信号通路的重要转录因子,在动物细胞增殖、分化、器官形成、垂体发育与胚胎发育等过程中发挥着重要作用。本研究以西农萨能奶山羊(Capra hircus)不同组织为实验材料,利用反转录PCR、TA克隆和生物信息学等方法克隆并分析PITX2基因序列,同时还利用实时荧光定量PCR(quantitative Real-time PCR,q RTPCR)技术研究该基因m RNA的表达规律。结果表明,奶山羊PITX2基因(Gen Bank登录号:KP721342)编码区全长978 bp,编码325个氨基酸,与牛(Bos taurus)、猪(Sus scrofa)、小鼠(Mus musculus)、大鼠(Rattus norvegicus)和人(Homo sapiens)的序列同源性分别为99%、95%、90%、90%和93%;PITX2基因在公、母羊的不同组织中均有不同程度表达,属广谱表达,其中,在公羊胰脏的表达量最高,小肠、肺和肌肉次之,在脾中的表达量最低,且胰、肺、小肠、肌肉的表达量均显著或极显著高于其他组织(P<0.05或P<0.01);在母羊肝脏中的表达量最高,乳腺和肺次之,脂肪中表达量最低,且乳腺组织的表达量显著高于心、肾、肌肉和脂肪等组织(P<0.05),提示该基因可能与泌乳性能有关。研究结果为进一步了解该基因在泌乳调控中的作用机制提供基础资料。Paired-liked homodomain transcription factor 2（PITX2） is an important transcription factor of Wnt/β- catenin signaling pathway, and plays an important role in cell proliferation, differentiation,organogenesis, pituitary development and embryonic development. Different tissues of Saanen dairy goats（Capra hircus） were collected as the experimental materials in this study. The PITX2 gene was cloned and analyzed using reverse transcription- polymerase chain reaction（RT- PCR）, TA cloning and bioinformatic methods, meanwhile the m RNA expression profiles of this gene was revealed using quantitative Real- timePCR（q RT- PCR）. The results showed that the coding sequence of PITX2 gene was 978 bp, encoding 325 amino acids, which shared 99%, 95%, 90%, 90% and 93% homology with Bos taurus, Sus scrofa, Mus musculus, Rattus norvegicus and Homo sapiens, respectively. The PITX2 gene expressed in varied tissues of rams and ewes, which showed a broad spectrum of gene expression. The expression of PITX2 gene was the highest in pancreas of ram, followed by small intestine, lungs and muscles, and the lowest in spleen.Moreover, the expression levels in pancreas, lungs, small intestine and muscles were significantly higher than that in other tissues. The expression of PITX2 gene was the highest in liver of ewe, followed closely by breast,lungs, and the lowest in adipose tissue. Moreover, the expression of breast was significantly higher than that in heart, kidney, muscle and adipose tissues, which suggested that this gene might be related with lactation performance. The results will lay a foundation for further study of the gene function in the regulation of lactation.

关 键 词：奶山羊 垂体同型框转录因子2基因(PITX2) 分子克隆 序列分析 m RNA表达 

分 类 号：S827[农业科学—畜牧学]