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酸核糖焦磷酸合成酶Ⅱ(phosphoribosylpyrophosphate synthetases 2,PRPS2),是生物体合成的关键酶,在控制肿瘤发生、促进细胞凋亡、抑制细胞增殖以及促进精子生成方面具有重要作用。为研究磷酸核糖焦磷酸合成酶Ⅱ基因(PRPS2)在猪(Sus scrofa)雄性不育方面的作用及基因编码的蛋白质功能,从Gen Bank下载猪及近缘物种的PRPS2 m RNA序列作为参考序列,设计特异引物扩增版纳微型猪近交系(Banna mini-pig inbred line,BMI)PRPS2基因,应用q RT-PCR分析15个重要组织的m RNA表达谱,并对蛋白质序列进行功能生物信息学分析。研究获得了BMI PRPS2 1 282 bp的c DNA序列(Gen Bank登录号:KU705637,对应的氨基酸登录号:AOC89055),编码318个氨基酸。组织表达谱显示,PRPS2基因在精囊腺中呈现高表达水平,在其他14个组织中呈现中低表达水平。生物信息学分析表明,PRPS2蛋白分子量(Mw)为34.83 k D,等电点(p I)为6.15,存在2个保守结构域,无跨膜结构,无信号肽序列,N末端和C末端疏水,有4类功能活性位点,位于细胞质的概率是89%;PRPS2基因在进化中高度保守。该研究结果将为进一步研究PRPS2基因在猪雄性不育方面的作用及功能提供参考。
Phosphoribosylpyrophosphate synthetases 2 (PRPS2), a key enzyme in biosynthesis, play an important role in controlling tumorigenesis, promoting apoptosis, inhibiting cell proliferation and promoting spermatogenesis. PRPS2 m RNA sequence of porcine and related species was downloaded as a reference sequence from GenBank to investigate the role of PRPS2 in male sterility of pig (Sus scrofa) and the function of the protein encoded by the gene. Primers were designed to amplify the PRPS2 gene of Banna mini-pig inbred line (BMI). QRT-PCR was used to analyze the m RNA expression profile of 15 important tissues and functional bioinformatics analysis of the protein sequence . The cDNA sequence of 1 282 bp of BMI PRPS2 was obtained (Gen Bank accession number: KU705637, the corresponding amino acid accession number: AOC89055), which encoded 318 amino acids. Tissue expression profiles showed that the PRPS2 gene was highly expressed in seminal vesicles, and showed low and intermediate expression levels in the other 14 tissues. Bioinformatics analysis showed that the PRPS2 protein has a molecular weight (Mw) of 34.83 kD and an isoelectric point (pi) of 6.15. There are two conserved domains, no transmembrane structure, no signal peptide sequence, N-terminal and C-terminal hydrophobic , There are 4 types of functional active sites, the probability of being located in the cytoplasm is 89%; PRPS2 gene is highly conserved in evolution. The results of this study will provide reference for further study on the role and function of PRPS2 gene in male sterility of pigs.