梅花鹿CSRP3基因克隆、生物信息学分析及其在肌肉组织中的表达分析

doi:10.16431/j.cnki.1671-7236.2025.02.024

摘要/Abstract

摘要： 【目的】探究梅花鹿富含半胱氨酸和甘氨酸蛋白3(cysteine and glycine-rich protein 3，CSRP3)基因的生物学作用及其在不同肌肉组织中的表达情况。【方法】以梅花鹿背最长肌为试验材料，克隆获得CSRP3基因，测序后进行相似性比对，构建系统进化树，并对CSRP3蛋白进行生物信息学分析。运用实时荧光定量PCR技术来研究梅花鹿背最长肌、臀大肌、股四头肌、肋间肌中CSRP3基因的表达差异。【结果】成功克隆出梅花鹿CSRP3基因，其编码区片段大小为585 bp,共编码194个氨基酸。BLAST比对结果显示，梅花鹿CSRP3基因与马鹿的相似性最高，为99.83%，与白尾鹿、牛、羊的相似性均超过95%，说明梅花鹿CSRP3基因在进化中具有高度保守性；根据梅花鹿CSRP3基因构建系统进化树，结果表明，梅花鹿与马鹿的亲缘关系最近，与虎和虎鲸的亲缘关系最远。CSRP3基因编码蛋白分子式为C₉₀₃H₁₄₀₃N₂₆₃O₂₇₅S₁₉，相对分子质量为20 952.81，理论等电点(pI)为8.89，脂肪系数为43.81，其二级结构以不规则卷曲为主；亚细胞定位于细胞核内。实时荧光定量PCR结果表明，CSRP3基因在梅花鹿不同部位肌肉组织中均有表达，在梅花鹿肋间肌中的表达量最高，与背最长肌、臀大肌、股四头肌存在显著性差异(P＜0.05)，而背最长肌、臀大肌和股四头肌之间CSRP3基因表达量差异并不显著(P＞0.05)。【结论】本研究成功克隆了梅花鹿CSRP3基因，其编码蛋白位于细胞核内； CSRP3基因在梅花鹿不同肌肉组织中存在差异表达。本研究结果为CSRP3基因在梅花鹿肉品质方面的调控机制研究提供了数据基础。

关键词: 梅花鹿; CSRP3基因; 克隆; 肌肉; 表达

Abstract: 【Objective】 The aim of this study was to investigate the biological function of cysteine and glycine-rich protein 3(CSRP3) gene in sika deer and its expression patterns in different muscle tissues. 【Method】 The longissimus dorsi muscle tissues of sika deer were used as experimental sample.The cDNA sequence of CSRP3 gene was cloned and sequenced,followed by sequence similarity alignment,construction of a phylogenetic tree,and bioinformatics analysis of CSRP3 protein. The expression differences of CSRP3 gene in longissimus dorsi muscle,gluteus maximus,intercostal muscle and quadriceps of sika deer was studied by Real-time quantitative PCR technology. 【Result】 The CSRP3 gene of sika deer was successfully cloned,and its coding region was 585 bp in size,encoding 194 amino acids.The BLAST comparison showed that CSRP3 gene of sika deer had the highest similarity with Cervus elaphus,at 99.83%,and the similarity with Odocoileus virginianus,Bos taurus,and Ovis aries were also over 95%,indicating that CSRP3 gene in sika deer had a high degree of conservation in evolution.The phylogenetic tree was constructed based on CSRP3 gene,and the results showed that sika deer was most closely related to that of Cervus elaphus and most distantly related to that of Panthera tigris and Orcinus orca.The molecular formula of CSRP3 gene-encoded protein was C₉₀₃H₁₄₀₃N₂₆₃O₂₇₅S₁₉,and the relative molecular mass was 20 952.81,the theoretical isoelectric point (pI) was 8.89,the fat coefficient was 43.81. The secondary structure of the protein was mainly random coil. Subcellular localization prediction indicated that the protein was located in the nucleus.Real-time quantitative PCR results showed that CSRP3 gene was expressed in different muscle tissues of sika deer.The expression level of this gene was the highest in the intercostal muscle of sika deer,with significant differences compared with the longissimus dorsi muscle,gluteus maximus,and quadriceps (P＜0.05),while the expression of CSRP3 gene in the longissimus dorsi muscle,gluteus maximus,and quadriceps were not significantly different (P＞0.05). 【Conclusion】 The CSRP3 gene of sika deer was successfully cloned in this study,which encoded a protein located in the nucleus of the cell.The CSRP3 gene was differentially expressed in different muscle tissues of sika deer.The results provided a data basis for investigating the regulatory mechanism of CSRP3 gene in sika deer meat quality.

Key words: sika deer; CSRP3 gene; clone; muscle; expression

中图分类号:

S825

王晴, 马瑜锴, 韩若冰, 邢海华, 孙子惠, 李和平. 梅花鹿CSRP3基因克隆、生物信息学分析及其在肌肉组织中的表达分析[J]. 中国畜牧兽医, 2025, 52(2): 740-748.

WANG Qing, MA Yukai, HAN Ruobing, XING Haihua, SUN Zihui, LI Heping. Cloning，Bioinformatics Analysis and Expression Analysis of CSRP3 Gene in Muscle Tissues of Sika Deer[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 740-748.

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