大足黑山羊卵泡抑素cDNA的克隆、序列分析及组织表达

Molecular Cloning,Sequence Analysis and Tissue Expression of Follistatin Gene in Dazu Black Goat

【目的】旨在克隆大足黑山羊卵泡抑素(follistatin)的基因序列,并研究其在不同组织中的表达差异。【方法】采用RT-PCR方法从大足黑山羊卵巢组织总RNA中克隆出follistatin的cDNA序列,用相关软件进行序列分析,并利用real-time PCR技术检测follistatin基因在不同组织中的表达。【结果】序列分析表明,大足黑山羊follistatin的cDNA序列全长为1 217 bp,包括该基因完整的开放阅读框(open reading frame,ORF)1 035 bp,编码344个氨基酸。与牛的核苷酸序列比对同源性高达98%。real-time PCR结果表明,follistatin基因能在心脏、肝脏、脾脏、肺、肾脏、卵巢和垂体中检测出来,在肾脏中表达量相对较高,在垂体中表达量较低。肾脏中follistatin mRNA含量显著高于垂体中follistatin mRNA含量(P<0.05);心脏中follistatin mRNA含量显著高于垂体中follistatin mRNA含量(P<0.05);其余各组织间follistatin mRNA含量差异均不显著(P>0.05)。【结论】成功克隆出大足黑山羊follistatin基因cDNA序列,该基因在肾脏中的表达高于其它组织。

【Objective】The objective of this experiment is to clone the sequence of Dazu black goat follistatin gene and explore the expression levels of follistatin gene in different tissues.【Method】The cDNA of follistatin gene was cloned from total RNA of ovary tissue in Dazu Black Goat by RT-PCR.The expression analysis of follistatin gene was predicted with some related software tools,and the expression of follistin gene in different tissues was analyzed with real-time PCR.【Result】Sequence analysis indicated that the cloned follistatin gene cDNA was 1 217 bp in length（HM138665） containing an entire open reading frame（ORF）,and encoding 344 amino acids.According to the Blast result in GenBank database,the cDNA nucleotides share 98% homology with Bos taurus.Real-time PCR analysis showed that the follistatin gene was expressed in spleen,lung,kidney,liver,heart,ovary and pituitary.The expression levels of follistatin gene were higher in kidney and lower in pituitary.The expression level of follistatin gene in kidney and heart were significantly higher than in pituitary（P0.05）.No significant difference was found between the other tissues（P0.05）.【Conclusion】The sequence of follistatin gene cDNA was obtained.And the follistatin mRNA expression was the hingest in kidney.