水牛浮舰蛋白2基因克隆及序列分析

Cloning and Sequence Analysis of Buffalo FLOT2 Gene

试验旨在克隆水牛浮舰蛋白2(Flotillin 2,FLOT2)基因并对其进行生物信息学分析,为揭示该基因在水牛发育繁殖中的作用奠定基础。本研究通过PCR扩增获得了FLOT2基因全长并进行克隆测序,结合生物信息学分析方法,预测及分析其蛋白质结构。结果表明,水牛FLOT2基因编码区长1 287bp,编码428个氨基酸,3′UTR区长277bp。多重序列比较分析显示,水牛FLOT2基因核苷酸序列与牛、山羊、绵羊、小鼠、马、猫和人相应序列的相似性分别为98%、98%、97%、90%、93%、92%和92%,系统进化树分析结果表明,FLOT2基因在不同物种以及进化的过程中具有高度保守性。对FLOT2蛋白质的分析表明,该蛋白呈弱酸性,无信号肽,细胞亚定位于细胞质中,存在SPFH_flotillin和SPFH_hflk等结构域。microRNA预测结果表明,bta-miR-2438、bta-miR-2379和bta-miR-1777a可能是其3′UTR潜在靶标。研究结果为今后阐明FLOT2基因在水牛繁殖性能中的功能,尤其是在配子及胚胎发生过程中的作用及分子机制奠定了基础。

In order to clarify the effect of Flotillin 2（FLOT2）gene on the reproductive performance of buffalo,buffalo FLOT2 gene was cloned and analyzed by bioinformatics.FLOT2 was cloned by PCR,cloned and sequenced,then its protein structure was predicted and analyzed by bioinformatics tools.The results showed that the coding region of buffalo FLOT2 gene was1 287 bp,3′UTR was 277 bp,encoded 428 amino acids.The buffalo FLOT2 gene shared 98%,98%,97%,90%,93%,92% and 92% of similar nucleotide sequence with that of Bos taurus,Ovis aries,Capra hircus,Mus musculus,Equus caballus,Felis catus and Homo sapiens,respectively.Phylogenetic tree analysis showed that FLOT2 gene was highly conserved in different species and evolution.FLOT2 protein was weakly acidic,without signal peptide,located in the cytoplasmic,and with the presence of SPFH_flotillin and SPFH_hflk domain.microRNA prediction showed bta-miR-2438,bta-miR-2379 and bta-miR-1777 amaybe target the 3′UTR of buffalo FLOT2.In a word,this study provided an important reference for surveying the regulation mechanism of FLOT2 gene in buffalo reproductive performance,especially,during buffalo gametogenesis and embryogenesis.