滩羊TCHH基因CDS区遗传结构及表达模式分析

doi:10.16431/j.cnki.1671-7236.2019.06.003

摘要/Abstract

摘要：

试验旨在分析滩羊TCHH基因的遗传结构，并检测该基因在同一月龄不同组织及不同月龄皮肤组织中的表达模式。通过基因同源扩增获得滩羊TCHH基因编码区（CDS）全长，并利用生物信息学技术对序列的同源性、蛋白基本性质及重复序列特征进行分析，同时利用半定量RT-PCR、实时荧光定量PCR对该基因在皮肤组织中的表达模式进行分析。结果显示，TCHH基因CDS区全长4 425 bp，包含两段重复序列，共编码1 474个氨基酸，滩羊TCHH氨基酸序列在物种内同源性较高，与山羊同源性可达96.3%，物种间的同源性较差，与人和家鼠的同源性分别为48.4%和40.5%。TCHH蛋白分子式为C₈₀₀₆H₁₃₁₈₇N₂₇₈₇O₂₆₆₁S₆，分子质量大小为191.26 ku，理论等电点为5.76，为亲水蛋白，其蛋白不稳定指数为119.37，稳定性较差。TCHH蛋白的氨基酸序列共包含2个保守区，分别为S-100型钙结合域和蛋白激酶结构域，富含α-螺旋，高达89.89%，推测其二级结构为长条棒状分子结构。组织表达谱结果发现，TCHH基因在皮肤组织中特异性表达，其mRNA表达量随着滩羊月龄的增加逐渐降低，与滩羊毛发卷曲性状的生长趋势一致，TCHH基因特殊的结构和表达模式表明该基因在滩羊的毛发卷曲性状中发挥着重要作用。本研究可为滩羊毛发生长的生物学机制研究提供重要的参考依据。

关键词: 滩羊; TCHH基因; 表达模式; 毛发卷曲

Abstract:

The aim of this experiment was to analyze the genetic structure of TCHH gene in Tan sheep and test the expression pattern of the gene in different tissues of the same month and skin tissue of different months.The full length of the coding sequence (CDS) region of TCHH gene was obtained by homologous cloning of genes,and the homology of the sequence,the basic properties of the protein and the characteristics of the repeat sequence were analyzed by bioinformatics technology.At the same time,the expression pattern of the gene in skin tissue was analyzed by semi-quantitative RT-PCR and Real-time PCR.The results showed that the TCHH gene CDS region was 4 425 bp in length and contained two repeats,encoding a total of 1 474 amino acids.The TCHH amino acid sequence of Tan sheep was highly homologous within the species,with 96.3% homology with Capra hircus,and the homology between species was poor.The homology with Homo sapiens and Mus musculus was 48.4% and 40.5%,respectively.The molecular formula of TCHH protein was C₈₀₀₆H₁₃₁₈₇N₂₇₈₇O₂₆₆₁S₆,the molecular mass was 191.26 ku,and the theoretical isoelectric point was 5.76,which was a hydrophilic protein.Its protein instability index was 119.37,and its stability was poor.The amino acid of this protein contained two conserved regions,S-100 calcium binding domain and protein kinase domain,which were rich in α-helix,up to 89.89%.It was speculated that the secondary structure was a long rod-like molecular structure.The results of tissue expression profiling showed that the gene was specifically expressed in skin tissue,and its mRNA expression decreased with the increase of the age of Tan sheep,which was consistent with the growth trend of the hair curling trait of Tan sheep wool.The specific structure and expression pattern of the TCHH gene indicated that this gene played an important role in the hair curling trait of Tan sheep.This study would provide important reference for the study of the biological mechanism of the occurrence of Tan sheep wool.

Key words: Tan sheep; TCHHgene; expression pattern; curly fleece

中图分类号:

S826

郑杰, 刘玉芳, 马露, 薛明. 滩羊TCHH基因CDS区遗传结构及表达模式分析[J]. 《中国畜牧兽医》, 2019, 46(6): 1585-1593.

ZHENG Jie, LIU Yufang, MA Lu, XUE Ming. Genetic Structure and Expression Pattern Analysis of TCHH in Tan Sheep[J]. , 2019, 46(6): 1585-1593.

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