猪卵泡抑素基因接拼形式克隆、表达及分析

Cloning,expression and bioinformatics analysis of two spliced forms of porcine follistatin gene

猪卵泡抑素基因(Follistatin,FS)具有调控动物生殖活动和肌肉生长的生物功能,但对猪的研究报道很少。本研究克隆了猪卵泡抑素基因两种接拼形式、比较分析其蛋白结构特征、进化关系和组织表达特性。研究结果表明,通过两对引物成功克隆了猪卵泡抑素基因的1045bp(FS-L)和964bp(FS-S)的两种接拼形式;与Genbank中的序列比对表明FS-L和FS-S的CDS序列与人、牛、家鼠、沟鼠、斑马鱼的卵泡抑素基因序列相似性在80%以上。对两种蛋白质结构域的比较发现,FS-S蛋白相比FS-L蛋白而言缺失了一个FS结构域。同源性分析表明,猪与牛的亲缘关系较近,与人、小鼠、沟鼠和斑马鱼的关系相对较远。组织表达分析表明,两种拼接形式在梅山猪不同组织中的表达存在差异,FS-L基因在梅山猪肌肉及生殖系统等组织中均有表达,而FS-S基因则主要在生殖系统中表达。表明FS-L在肌肉生长中发挥作用,而FS-S主要调控动物的生殖功能。

Porcine follistatin gene （ Follistatin, FS） plays roles in regulation of animal reproductive activities and muscle growth, however very few studies on the swine FS are available. In this study, two spliced forms of porcine FS gene were cloned, and the domain and protein structure, evolutionary relationship and tissue expression pattern were investigated. The results showed that two spliced forms （ 1 045bp, FS-L and 964bp, FS-S） were cloned suc- cessfully with tow pairs of primers; the sequences were aligned with sequences of human, mouse, rat, and ze- brafish deposited in Genbank and the identity was higher than 80%. Based on the comparison of the two protein do- mains, FS-S protein is missing one FS domain compared to FS-L protein. The homology analysis showed that pigs and cattle share closer relationship but the relationships between pig and human, mouse, rat, and zebra fish are relatively distant. The tissue expression analysis showed that the two splice forms display different patterns among Meishan pig tissues, the FS-L form was expressed in muscle and reproductive system and others, but FS-S form was mainly expressed in reproductive system. It implies that FS-L may play a role in muscle growth, while the FS-S may mainly regulates animal reproductive function.