版纳微型猪近交系ASB17基因克隆及睾丸组织特异表达研究

发布时间：2018-04-19 00:12

本文选题：ANK(锚定蛋白重复) + SOCS(细胞因子信号抑制的蛋白)　；参考：《河北农业大学学报》2017年05期

【摘要】：ASB17基因与睾丸形成和精子发生密切相关。为研究ASB17基因在猪睾丸功能及精子发生过程中的作用,本研究从GenBank下载猪及近缘物种ASB17序列作为参考序列,设计特异引物从版纳微型猪近交系(BMI)睾丸组织中扩增ASB17基因完全编码序列及部分侧翼序列,对ASB17翻译的蛋白质序列进行多种功能生物信息学分析并构建多物种系统进化树,最后用实时荧光定量PCR技术检测BMI 15个重要组织的ASB17 mRNA转录表达水平。结果表明,扩增得到BMI ASB17编码区序列长888bp,编码295个氨基酸。功能生物信息学分析表明ASB17存在2个保守结构域ANK和SOCS_box,无跨膜螺旋结构,无N端信号肽序列,二级结构以α螺旋为主,N末端和C末端均亲水,有4类功能活性位点,位于细胞质的概率70.6%。系统进化分析表明,ASB17基因在进化中高度保守,且与牛、羊的亲缘关系最近,符合物种的系统分类学。多组织相对荧光定量表达分析表明ASB17基因在睾丸中特异高表达,在心、肝等其他14个组织中不表达。研究结果为探索BMI ASB17基因在睾丸形成和精子发生过程中的作用及功能奠定基础。
[Abstract]:ASB17 gene is closely related to testicular formation and spermatogenesis.In order to study the role of ASB17 gene in testis function and spermatogenesis of pigs, we downloaded the ASB17 sequences of porcine and related species from GenBank as reference sequences.A specific primer was designed to amplify the complete coding sequence and partial flanking sequence of the ASB17 gene from the testis of BMI, a miniature pig inbred strain of Banna, and to analyze the protein sequence translated by ASB17 in a variety of functional bioinformatics and to construct a multispecies phylogenetic tree.Finally, ASB17 mRNA transcriptional expression in 15 important tissues of BMI was detected by real-time fluorescence quantitative PCR.The results showed that the sequence of BMI ASB17 coding region was 888bp, encoding 295 amino acids.Functional bioinformatics analysis showed that ASB17 had two conserved domains, ANK and SOCs, no transmembrane helical structure and no N-terminal signal peptide sequence. The secondary structure consisted of 伪 -helix and hydrophilic at both N-terminal and C-terminal, with four functional active sites.The probability of being located in the cytoplasm was 70.6.Phylogenetic analysis showed that the ASB17 gene was highly conserved in evolution and had the closest phylogenetic relationship with cattle and sheep, which was consistent with the phylogeny of the species.Quantitative analysis of multitissue relative fluorescence showed that ASB17 gene was specifically overexpressed in testis, but not expressed in 14 other tissues, such as heart, liver and so on.The results laid a foundation for exploring the role and function of BMI ASB17 gene in testicular formation and spermatogenesis.
【作者单位】：云南农业职业技术学院;云南省农业科学院粮食作物研究所;云南农业大学云南省版纳微型猪近交系重点实验室;云南农业大学动物科学技术学院;
【基金】：云南省教育厅科学研究基金项目(2017ZZX099) 国家自然科学基金项目(31660637;31460580;31660650)
【分类号】：S828.2

【相似文献】