陕北白绒山羊MSX2基因克隆及表达分析

Cloning and Expression of MSX2 Gene in Shanbei White Cashmere Goat

本研究旨在克隆陕北白绒山羊MSX2基因cDNA全序列,并对其序列特征及其表达规律进行分析。试验以陕北白绒山羊皮肤组织为材料,采用RT-PCR技术,对陕北白绒山羊MSX2基因的cDNA序列进行克隆,并进行生物信息学分析。采用实时荧光定量PCR检测并分析MSX2mRNA在陕北白绒山羊毛囊不同发育时期皮肤组织中的表达规律。结果表明,山羊MSX2基因cDNA全长804bp(GenBank登录号:JQ617290),为一个完整的开放阅读框(ORF),编码267个氨基酸,与牛(NM_001079614)、人(NM_002449)、犬(NM_001003098)、黑猩猩(NM_001135625)、大鼠(NM_012982)和小鼠(NM_013601)的相似性分别为96%、90%、89%、88%、87%和85%;MSX2基因在4月和11月的相对表达量出现峰值,与其他月份差异显著(P<0.05)。山羊MSX2基因编码区核苷酸序列高度保守。MSX2基因在陕北白绒山羊皮肤毛囊发育的4月份和11份月高度表达,其余月份均有不同程度的表达,推测MSX2基因对毛囊周期调控具有一定的作用。

This experiment was conducted to obtain and analyze cDNA sequence of goat MSX2,and to analyze the sequence character and expression rule.By reverse transcription PCR,the full-length cDNA of MSX2 was cloned from skin of Shanbei White Cashmere goat,and the expression of MSX2 in different development periods of follicle were examined by quantitative real-time PCR.The results showed that the cDNA of goat MSX2 contained 804 nucleotides,including a complete open reading frame（ORF） encoding 267 amino acids.The amino acid sequence shared 96%,90%,89%,88%,87% and 85% identity with those of Bos taurus,Homo sapiens,Canis familiaris,Pan troglodytes,Rattus norvegicus,Mus musculus,respectively.The result of real-time quantitative PCR showed that MSX2 were higher expressed in April and November than that in other months（P〈0.05）.The amino acid sequences of MSX2 were highly conservative.MSX2 was highly expressed at April and November,and it was expressed in different extent in the other months,which indicated that MSX2 gene might be involved in molecular regulation of hair growth cycle.