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

Abstract

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

CLC Number:

S825

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.

References

[1] 翟晶,左璐雅,付晶，等.双阳梅花鹿鹿肉营养成分分析[J].内蒙古农业大学学报(自然科学版),2016,37(6):128-132.ZHAI J,ZOU L Y,FU J,et al.Analysis of nutrient component in muscle of Shuangyang sika deer [J].Journal of Inner Mongolia Agricultural University(Natural Science Edition),2016,37(6):128-132. (in Chinese)
[2] HOFFMAN L C,WIKLUND E.Game and venison-meat for the modern consumer [J].Meat Science,2006,74(1):197-208.
[3] JIMENEZ-COLMENERO F,CARBALLO J,COFRADES S.Healthier meat and meat products:Their role as functional foods [J].Meat Science,2001,59(1):5-13.
[4] LIEBHABER S A,EMERY J G,URBANEK M,et al.Characterization of a human cDNA encoding a widely expressed and highly conserved cysteine-rich protein with an unusual zinc-finger motif [J].Nucleic Acids Research,1990,18(13):3871-3879.
[5] WEISKIRCHEN R,PINO J D,MACALMA T,et al.The cysteine-rich protein family of highly related LIM domain proteins [J].The Journal of Biological Chemistry,1995,270(48):28946-28954.
[6] ARBER S,CARONI P.Specificity of single LIM motifs in targeting and LIM/LIM interactions in situ [J].Genes & Development,1996,10(3):289-300.
[7] TARRO G.Tumor liberated protein (TLP):Its potential for diagnosis and therapy [J].Anticancer Research,1999,19(3A):1755-1757.
[8] LOUIS H A,PINO J D,SCHMEICHEL K L,et al.Comparison of three members of the cysteine-rich protein family reveals functional conservation and divergent patterns of gene expression [J].The Journal of Biological Chemistry,1997,272(43):27484-27491.
[9] VAFIADAKI E,ARVANITIS D A,PAPALOUKA V,et al.Muscle lim protein isoform negatively regulates striated muscle actin dynamics and differentiation [J].FEBS Journal,2014,281(14):3261-3279.
[10] ARBER S,HUNTER J J,ROSS J,et al.MLP-deficient mice exhibit a disruption of cardiac cytoarchitectural organization,dilated cardiomyopathy,and heart failure [J].Cell,1997,88(3):393-403.
[11] GEIER C,GEHMLICH K,EHLER E,et al.Beyond the sarcomere CSRP3 mutations cause hypertrophic cardiomyopathy [J].Human Molecular Genetics,2008,17(18):2753-2765.
[12] HAN S,CUI C,WANG Y,et al.Knockdown of CSRP3 inhibits differentiation of chicken satellite cells by promoting TGF-β/Smad3 signaling [J].Gene,2019,707:36-43.
[13] SHAN Y J,JI G G,MING Z,et al.Use of transcriptome sequencing to explore the effect of CSRP3 on chicken myoblasts [J].Journal of Integrative Agriculture,2023,22(4):1159-1171.
[14] HE H,ZHANG H L,LI Z X,et al.Expression,SNV identification,linkage disequilibrium,and combined genotype association analysis of the muscle-specific gene CSRP3 in Chinese cattle [J].Gene,2014,535(1):17-23.
[15] 彭章蓉,朱勇臻,王新新，等.鹿肉品质及影响因素研究进展[J].特产研究,2022,44(6):136-140.PENG Z L,ZHU Y Z,WANG X X,et al.Current status of venison research and progress of research on factors influencing venison quality[J].Special Wild Economic Animal and Plant Research,2022,44(6):136-140.(in Chinese)
[16] ROTZOLL N,DUNKEL A,HOFMANN T.Quantitative studies,taste reconstitution,and omission experiments on the key taste compounds in Morel mushrooms (Morchella deliciosa fr.) [J].Journal of Agricultural and Food Chemistry,2006,54(7):2705-2711.
[17] 白佳灵,王会,柴志欣，等.牦牛CSRP3基因的克隆及组织表达分析[J].生物技术通报,2020,36(6):200-207.BAI J L,WANG H,CHAI Z X,et al.Cloning and tissue expression analysis of CSRP3 gene in yak [J].Biotechnology Bulletin,2020,36(6):200-207.(in Chinese)
[18] 陈云,潘鹏丞,陈钊，等.陆川猪CSRP3基因克隆及其组织表达差异分析[J].南方农业学报，2022,53(4):977-984.CHEN Y,PAN P C,CHEN Z,et al.Cloning and differential expression analysis of CSRP3 gene in Luchuan pig [J].Journal of Southern Agriculture,2022,53(6):977-984.(in Chinese)
[19] VAFIADAKI E,ARVANITIS D A,SANOUDOU D.Muscle LIM protein:Master regulator of cardiac and skeletal muscle functions [J].Gene,2015,566(1):1-7.
[20] CHANG Y,GENG F,HU Y,et al.Zebrafish cysteine and glycine-rich protein 3 is essential for mechanical stability in skeletal muscles [J].Biochemical and Biophysical Research Communications,2019,511(3):604-611.
[21] 单艳菊,姬改革,章明，等. CSRP3基因在鸡胚胎期胸肌、成肌细胞中的表达及其与胸肌质量的相关性研究[J].中国畜牧杂志,2023,59(1):131-135.SHAN Y J,JI G G,ZHANG M,et al.The expression of CSRP3 gene in pectoral muscle and myoblasts in chicken embryonic stage and its correlation with pectoral muscle quality [J].Chinese Journal of Animal Science,2023,59(1):131-135.(in Chinese)
[22] PRADERI L A.Hemangioma of the intercostal muscle [J].Boletin Sociedad de Cirugia del Uruguay,1960,31:131-136.

Cloning，Bioinformatics Analysis and Expression Analysis of CSRP3 Gene in Muscle Tissues of Sika Deer

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	AI Xiaonan, CHENG Lifen, BAI Pu, CHEN Zhenghao, XUE Lina, ZHOU Shenghua. Cloning and Bioinformatics Analysis of ACSL1 Gene and Its Expression Pattern During Adipocyte Differentiation in Lingqiu Qingbei Goats [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 499-511.
[2]	SHEN Yangyang, AN Zhenjiang, GAO Linna, XIA Shuwen, CHEN Kunlin, DING Qiang, GAO Yundong, XUE Jinguo, ZHONG Jifeng, WANG Huili. Comparative Analysis of Muscle Transcriptome of Xuzhou Cattle with Different Tenderness [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 522-533.
[3]	LEI Zhigang, PAN Hong, ZHANG Dandan, LIU Quanhui, SUN Zhe, DENG Shan, HUANG Ben. Cloning, Bioinformatics Analysis and Eukaryotic Expression Vector Construction of MAPK8 Gene in Mashan Black Goats [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 534-544.
[4]	LI Jingxuan, LIN Yanjiao, HUANG Qiongjun, HAN Xinyan, ZHANG Yuelang. Research Progress on Non-coding RNA Related to Skeletal Muscle Development in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 582-592.
[5]	XIE Beiyiting, WANG Yue, MENG Chunhua, QIAN Yong, ZHANG Jun, ZHANG Jianli, WANG Huili, CAO Shaoxian, LI Yinxia. Cloning and Sequence Analysis of PSMB9 Gene in Hu Sheep and Its Effect on Myoblast Proliferation [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 1-12.
[6]	HUANG Yao, YAO Naiquan, YUAN Weitao, MAO Aipeng, ZHANG Ting, XU Chao. Isolation and Identification of Candida rugosa DZ018 from Deer and Its Probiotic Properties in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 389-399.
[7]	HAN Fengye, LIU Ying, PAN Chenfan, ZHANG Qianyi, CHEN Xiaoyun, ZHU Zhen, YIN Chunsheng, WEN Yongjun, WANG Fengxue, DU Jige. Expression and Preparation of Monoclonal Antibodies of Clostridium perfringens Alpha-Toxin Mutant [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 442-450.
[8]	ZHOU Fangting. Cloning,Molecular Characteristics and Tissue Differential Expression Analysis of KLF15 Gene in Buffalo [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 3715-3725.
[9]	SUN Huimin, FAN Bingfeng, ZHANG Xulin, SHAO Jing, LIU Lixiang, XU Baozeng. Transcriptomic Analysis of Blood in Early Pregnancy at Different Time Points in Sika Deer [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 3892-3908.
[10]	LI Shunan, LIU Dapeng, LI Zhinan, LI Yating, ZHANG Beibei, HOU Shuisheng, ZHOU Zhengkui. Study on the Differences in Meat Quality and Sensory Evaluation of Leg Muscle Between Pekin Ducks and Liancheng White Ducks [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 3939-3947.
[11]	GUO Chenyun, SONG Jinxing, WANG Mengxiang, SUN Junru, YAN Ruoqian, XIE Caihua, WU Yanan, ZHANG Gaiping. Prokaryotic Expression,Purification and Polyclonal Antibodies Preparation of African Swine Fever Virus B602L Protein [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 3980-3989.
[12]	CHENG Jing, CHEN Peilin, GUO Yu, CUI Jinqiang, JIANG Bo, ZHOU Linyi, LIU Wenxiao, LI Huanrong, LI Yongqing. Prokaryotic Expression and Monoclonal Antibody Preparation of Bovine Promyelocytic Leukemia Protein [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 4014-4024.
[13]	CUI Yanyan, WANG Guan, HAO Junfang, LI Zhiqiang, YU Fuchang, SHI Ke, XI Li, QI Meng. Prokaryotic Expression of Anaplasma phagocytophilum MSP4 Protein and Preparation of Its Polyclonal Antibodies [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 4036-4042.
[14]	XU Haotian, YU Yuetong, LI Jing, MA Zhiyuan, YANG Bin, WANG Zekun, TUO Haixin, QI Meng. Comparative Analysis of miRNA Expression Profiles in Bovine Mammary Epithelial Cells Treated with Lipopolysaccharide and Calcium Ions [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 4066-4079.
[15]	LI Rong, FU Guowen, ZENG Qin, WU Jiao, YUAN Jiarui, LIU Yabo, SILANG Yuzhen, FAN Yueyuan. Analysis of the Differences of Chemokine-binding Protein Gene Between Ovine Contagious Pustular Dermatitis Virus Oral and Visceral Infection Strains and Their Prokaryotic Expression [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(8): 3506-3518.