版纳微型猪近交系FANK1基因CDS克

发布时间：2018-04-13 18:08

本文选题：纤连蛋白(FN) + 锚蛋白重复(ANK)　；参考：《四川农业大学学报》2017年03期

【摘要】：【目的】获得猪FANK1基因CDS序列、组织表达和蛋白质功能信息。【方法】以GenBank下载的猪及近缘物种的FANK1 mRNA序列为参考序列,设计特异引物从版纳微型猪近交系(BMI)睾丸组织中扩增FANK1基因完全编码序列及部分侧翼序列。应用实时荧光定量PCR技术检测BMI 15个重要组织的FANK1 mRNA转录表达水平,并对FANK1翻译的蛋白质序列进行多种功能生物信息学分析,预测FANK1蛋白质的功能,最后构建FANK1多物种氨基酸系统进化树。【结果】获得了BMI FANK1编码区序列长1 041 bp,编码346个氨基酸,序列已提交GenBank,基因登录号为KU705617和KU705618,对应的氨基酸登录号为AOC89035和AOC89036。基因组结构分析表明FANK1基因定位于猪14号染色体,有11个外显子和10个内含子。多组织相对荧光定量表达分析表明FANK1基因在睾丸、尿道球腺和精囊腺中呈高表达水平;在其他组织中呈中低表达水平。功能生物信息学分析表明FANK1蛋白质含有2种保守结构域FN3和ANK,无跨膜螺旋结构,无N端信号肽序列,二级结构以无规卷曲为主,N末端和C末端均亲水,有4类功能活性位点。系统进化分析表明,FANK1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。【结论】成功克隆了版纳微型猪近交系FANK1基因的CDS序列并进行了多种组织表达分析,为后续研究FANK1基因在小型猪精细胞分裂及调节信号通路方面的作用及功能奠定基础。
[Abstract]:[objective] to obtain the CDS sequence, tissue expression and protein function information of porcine FANK1 gene. [methods] the FANK1 mRNA sequence of porcine and related species downloaded by GenBank was used as reference sequence.A specific primer was designed to amplify the complete coding sequence and partial flanking sequence of FANK1 gene from BMI testis of Banna miniature pig inbred line.The transcription and expression of FANK1 mRNA in 15 important tissues of BMI were detected by real-time fluorescence quantitative PCR, and the protein sequences translated by FANK1 were analyzed by multifunctional bioinformatics to predict the function of FANK1 protein.Finally, the phylogenetic tree of FANK1 multi-species amino acids was constructed. [results] the coding region of BMI FANK1 was 1 041 BP, encoding 346 amino acids. The sequence was submitted to GenBank, the gene accession numbers were KU705617 and KU705618, and the corresponding amino acid login numbers were AOC89035 and AOC89036.Genomic structure analysis showed that FANK1 gene was located on pig chromosome 14 with 11 exons and 10 introns.Quantitative analysis of relative fluorescence in multiple tissues showed that the expression of FANK1 gene was high in testis urethra glandular gland and seminal vesicle gland and low expression level in other tissues.Functional bioinformatics analysis showed that the FANK1 protein contained two conserved domains, FN3 and ANK, no transmembrane helical structure, no N-terminal signal peptide sequence, and the secondary structure was mainly composed of random curls, which were hydrophilic at the N-terminal and C-terminal, and had four functional active sites.Phylogenetic analysis showed that the FANK1 gene was highly conserved in evolution and had a close relationship with cattle and sheep.[conclusion] the CDS sequence of the FANK1 gene of Banna miniature pig inbred line was cloned successfully and the expression of various tissues was analyzed.The results provide a basis for further study on the role and function of FANK1 gene in spermatocyte division and regulation of signaling pathway in miniature pigs.
【作者单位】：华中农业大学生命科学技术学院;云南农业大学动物科学技术学院;
【基金】：国家自然科学基金项目(31660637;31460580;31660650)
【分类号】：Q78;S828

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