猪睾丸表达序列37(TEX37)克隆、mRNA表达及蛋白质功能生物信息学分析  

Cloning,mRNA Expression and Protein Bioinformatics Analysis of the Expressed Sequence 37(TEX37)of Porcine Testis

作　　者：霍海龙[1] 张霞[2] 范俐 赵筱 王世雄[1] 王荣琼[1] 刘丽仙[1] 霍金龙[2] HUO Hailong;ZHANG Xia;FAN Li;ZHAO Xiao;WANG Shixiong;WANG Rongqiong;LIU Lixian;HUO Jinlong(Yunnan Vocational and Technical College of Agriculture,Kunming 650031,China;Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Institute of Food Crops,Yunnan Academy of Agricultural Sciences,Kunming 650205,China)

机构地区：[1]云南农业职业技术学院,云南昆明650031 [2]云南农业大学动物科学技术学院,云南昆明650201 [3]云南省农业科学院粮食作物研究所,云南昆明650205

出　　处：《云南农业大学学报（自然科学版）》2020年第5期775-780,共6页Journal of Yunnan Agricultural University:Natural Science

基　　金：云南省教育厅科学研究基金项目(2018JS582);国家自然科学基金项目(31660637;31460580;31660650)。

摘　　要：【目的】以版纳微型猪近交系(Banna mini-pig inbred line,BMI)为材料,初步探索猪睾丸表达序列37(TEX37)基因的特性和功能,为该基因在猪精子生成过程的作用研究奠定基础。【方法】利用Oligo7软件设计特异引物扩增BMI TEX37基因整个编码区,应用实时荧光定量PCR技术检测BMI 15个组织的TEX37 mRNA表达谱,并初步预测TEX37蛋白质的功能,最后构建TEX37的多物种氨基酸系统进化树。【结果】扩增得到BMI TEX37编码区序列长561 bp,编码186个氨基酸,基因登录号KU705619,氨基酸登录号AOC89037。多组织相对荧光定量表达分析表明:TEX37基因在睾丸中呈相对最高表达,在其他组织中呈相对较低表达。生物信息学分析表明:TEX37蛋白质含有TSC21结构域,二级结构以无规则卷曲为主,N末端疏水,C末端亲水,有2类磷酸化位点,无跨膜螺旋结构和信号肽。系统聚类表明:BMI与其他物种相似度在67%以上,进化较保守。【结论】该研究可为TEX37基因在猪精子生成方面的作用机理研究提供参考。[Purpose]The aim of this study was to obtain the properties and function of testis expression sequence 37(TEX37)gene in Banna mini-pig inbred line(BMI)for further study of the gene in spermatogenesis role.[Method]The specific primers were designed by using Oligo7 and TEX37 complete coding sequence was amplified from BMI.The TEX37 expression profiles of mRNA in 15 tissues of BMI was detected by real-time quantitative PCR.TEX37 protein sequence was conducted functional analysis and established phylogenetic tree.[ Result] We obtained a 561 bp coding sequenceof BMI TEX37 gene with GenBank accession number of KU705619, which encodes a proteinof 186 amino acids with GenBank accession number AOC89037. real-time quantitative PCR in multitissuesindicated that TEX37 gene has the highest expressed in the BMI testis tissue and low expressionin the other tissues. Further functional bioinformatics analysis revealed TEX37 protein containeda conserved domains TSC21 without transmembrane helix and signal peptide. Its secondary structureis predominantly random coils and its N-terminus hydrophobic and C-terminal hydrophilic. TheTEX37 protein has two kinds of phosphorylation sites. Phylogenetic analysis indicated that BMI hadthe over 67% genetic similarity with other species.[Conclusion]This study will accumulate somebasic data for further research of the TEX37 gene function in pig spermatogenesis.

关 键 词：睾丸表达序列37(TEX37) 版纳微型猪近交系 精子生成 荧光定量PCR 生物信息学 

分 类 号：S828.2[农业科学—畜牧学]