牦牛CSRP3基因的克隆及组织表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0873

生物技术通报 ›› 2020, Vol. 36 ›› Issue (6): 200-207.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0873

牦牛CSRP3基因的克隆及组织表达分析

白佳灵^1,2, 王会^1,2, 柴志欣^1,2, 王吉坤^1,2, 王嘉博^1,2, 武志娟^1,2, 信金伟³, 钟金城^1,2, 陈智华², 姬秋梅³

1. 西南民族大学青藏高原动物遗传资源保护与利用四川省教育部重点实验室,成都 610041;
2. 西南民族大学青藏高原研究院,成都 610041;
3. 省部共建青稞和牦牛种质资源与遗传改良国家重点实验室,拉萨 850000

收稿日期:2019-09-19 出版日期:2020-06-26 发布日期:2020-06-28
作者简介:白佳灵,硕士研究生,研究方向：分子生态学;E-mail：929712733@qq.com
基金资助:
西南民族大学研究生创新型科研项目（CX2019SZ97）,国家肉牛牦牛产业技术体系项目（CARS-37）,西藏自治区财政专项项目（XZNKY-2019-C-052）,青藏高原生态畜牧业协同创新中心开放基金（QZGYXT02）

Cloning and Tissue Expression Analysis of CSRP3 in Yak

BAI Jia-ling^1,2, WANG Hui^1,2, CHAI Zhi-xin^1,2, WANG Ji-kun^1,2, WANG Jia-bo^1,2, WU Zhi-juan^1,2, XIN Jin-wei³, ZHONG Jin-cheng^1,2, CHEN Zhi-hua², JI Qiu-mei³

1. Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Sichuan Province and Ministry of Education,Southwest Minzu University,Chengdu 610041;
2. Institute of Tibetan Plateau Research,Southwest Minzu University,Chengdu, 610041;
3. State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement,Lhasa 850000

Received:2019-09-19 Published:2020-06-26 Online:2020-06-28

摘要/Abstract

摘要： CSRP3基因（Cysteine and glycine-rich protein 3,CSRP3）编码CRP3蛋白,是一个肌发生的正调节因子,可通过多种方式在肌肉发育和肌肉细胞结构维持中起重要作用。通过对牦牛CSRP3基因进行克隆及组织表达谱分析,为后续提高牦牛肉品质的研究提供基础数据。采用RT-PCR方法克隆牦牛CSRP3基因CDS区;再对其进行序列分析及蛋白结构和功能预测等生物信息学分析;最后利用实时荧光定量PCR技术检测该基因在牦牛不同组织中的表达量。牦牛CSRP3基因CDS区长585 bp,编码194个氨基酸;CSRP3基因的系统进化树结果显示,牦牛与黄牛的亲缘性最近,其次是绵羊。牦牛CSRP3基因编码的蛋白为偏碱性不稳定亲水蛋白,无跨膜结构和信号肽,为非分泌蛋白,含有磷酸化位点22个,N-糖基化位点2个,O-糖基化位点7个;存在两个LIM结构域,属于LIM结构域蛋白质超家族成员,主要分布于细胞核中;二级结构以无规卷曲为主,三级结构的最佳模型为1b8t.1.A;实时荧光定量PCR结果显示,牦牛CSRP3基因在臀大肌中有较高表达量。生物信息学分析结果显示,CRP3蛋白含有两个LIM结构域,主要分布在细胞核中,实时荧光定量PCR结果显示牦牛CSRP3基因在臀大肌中具有较高表达量,为牦牛CSRP3基因在牦牛肉品质方面的调控机制研究提供了基础数据。

关键词: 牦牛, CSRP3基因, 组织表达, 生物信息学分析

Abstract: The CSRP3 gene（cysteine and glycine-rich protein 3）encodes CRP3 protein,which is a positive regulator of myogenesis and plays an important role in muscle development and structural maintenance of muscle cells. Through cloning and tissue expression profile analysis of yak CSRP3 gene,the basic data may be provided for the further study of improving yak meat quality. The CDS region of yak CSRP3 gene was cloned by RT-PCR and analyzed by bioinformatics of sequence analysis,protein structure and function prediction. Finally,the expression of the gene in yak tissues was detected by qPCR. The results showed that the CDS region length of CSRP3 gene in yak was 585 bp,and a total of 194 amino acids were encoded. The phylogenetic tree demonstrated that yak had the closest relationship with cattle,followed by sheep. The protein encoded by CSRP3 gene was an alkaline unstable hydrophilic protein without transmembrane structure and signal peptide,not a secretory protein. There were 22 phosphorylation sites,2 N-glycosylation sites,7 O-glycosylation sites and 2 LIM domains,and it belonged to the superfamily member of LIM structure domain protein. The protein was mainly distributed in the nucleus. The secondary structure was mainly composed of random coils. The best model of the tertiary structure was 1b8t.1.A. The bioinformatics analysis revealed that the CRP3 protein contained 2 LIM domains and mainly distributed in the nucleus. The qPCR results showed that the yak CSRP3 gene had higher expression in the gluteus maximus. These results provide basic data for the research on whether the CSRP3 gene may improve the quality of yak meat.

Key words: Yak, CSRP3 gene, tissue expression, bioinformatics analysis

白佳灵, 王会, 柴志欣, 王吉坤, 王嘉博, 武志娟, 信金伟, 钟金城, 陈智华, 姬秋梅. 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 Jia-ling, WANG Hui, CHAI Zhi-xin, WANG Ji-kun, WANG Jia-bo, WU Zhi-juan, XIN Jin-wei, ZHONG Jin-cheng, CHEN Zhi-hua, JI Qiu-mei. Cloning and Tissue Expression Analysis of CSRP3 in Yak[J]. Biotechnology Bulletin, 2020, 36(6): 200-207.

参考文献

[1] Guanqing L, Chunlan Z, Guizhi W, et al.Molecular cloning, characterization and tissue specificity of the expression of the ovine CSRP2 and CSRP3 genes from Small-tail Han sheep(Ovis aries)[J]. Gene, 2016, 580(1):47-57.
[2] Arber S, Halder G, Caroni P.Muscle LIM protein, a novel essential regulator of myogenesis, promotes myogenic differentiation[J]. Cell, 1994, 79(2):221-231.
[3] Barash I A, Mathew L, Lahey M, et al.Muscle LIM protein plays both structural and functional roles in skeletal muscle[J]. AJP Cell Physiology, 2005, 289(5):C1312-C1320.
[4] Knöll R, Kostin S, Klede S, et al.A common MLP(muscle LIM protein)variant is associated with cardiomyopathy[J]. Circulation Research, 2010, 106(4):695-704.
[5] Fatkin D, Graham RM.Molecular Mechanisms of Inherited Cardiomyopathies[J]. Physiological Reviews, 2002, 82(4):945-980.
[6] 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.
[7] Buyandelger B, Ng K E, Miocic S, et al.MLP(muscle LIM protein)as a stress sensor in the heart[J]. Pflügers Archiv - European Journal of Physiology, 2011, 462(1):135-142.
[8] Rashid M M, Runci A, Polletta L, et al.Muscle LIM protein/CSRP3:a mechanosensor with a role in autophagy[J]. Cell Death Discovery, 2015, 1:15014.
[9] Arvanitis D A, Vafiadaki E, Papalouka V, et al.Muscle Lim Protein and Myosin Binding Protein C form a complex regulating muscle differentiation[J]. Biochimica et biophysica acta, 2017, 1864(12):2308-2321.
[10] Carole O, Andreia G, Valérie N, et al.CSRP3 mediates polyphenols-
induced cardioprotection in hypertension[J]. The Journal of Nutritional Biochemistry, 2019, 66:29-42.
[11] Xu X, Qiu H, Du Z Q, et al.Porcine CSRP3:polymorphism and association analyses with meat quality traits and comparative analyses with CSRP1 and CSRP2[J]. Molecular Biology Reports, 2010, 37(1):451-459.
[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] He H, Zhang H L, Li ZX, 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.
[14] 张容昶. 中国的牦牛[M]. 兰州:甘肃科学技术出版社, 1989.
[15] 蔡立. 中国的牦牛[J]. 西南民族大学学报:自然科学版, 1980(2):1-5, 7-12.
[16] 信金伟, 张成福, 姬秋梅, 等. 类乌齐牦牛产肉性能及肉品质分析[J]. 湖北农业科学, 2017, 56(3):501-505.
[17] 孙寿永, 张浩. 海门山羊不同部位肌肉中氨基酸含量的研究[J]. 中国畜牧兽医, 2012, 39(12):77-81.
[18] 王一冰, 蒋守群, 苟钟勇, 等. 赖氨酸的生物学功能及肉鸡营养需要量的研究[J]. 广东畜牧兽医科技, 2018, 43(6):17-20.
[19] 王丽慧. 日粮不同水平半胱氨酸对滩寒杂交羊日增重、屠宰率与肉品质的影响[D]. 杨凌:西北农林科技大学, 2015.
[20] 刘冠卿. 小尾寒羊CSRP2、CSRP3和FSTL1基因克隆、结构特征及组织表达分析[D]. 泰安:山东农业大学, 2015.
[21] Vafiadaki E, Arvanitis DA, Sanoudou D.Muscle LIM Protein:Master regulator of cardiac and skeletal muscle functions[J]. Gene, 2015, 566(1):1-7.
[22] 孙向阳, 刘书宁, 侯照远, 等. LIM蛋白Ajuba的研究进展[J]. 药物生物技术, 2017, 24(3):263-268.
[23] 俞红贤, 莫重存. 青海牦牛心脏的比较解剖学特点[J]. 黑龙江畜牧兽医, 2000(6):4-5.
[24] Ehsan M, Jiang H, Thomson KL, et al.When signalling goes wrong:pathogenic variants in structural and signalling proteins causing cardiomyopathies[J]. Journal of Muscle Research and Cell Motility, 2017, 38(Pt 9):303-316.
[25] Willmann R, Kusch J, Sultan K R, et al.Muscle LIM protein is upregulated in fast skeletal muscle during transition toward slower phenotypes[J]. AJP Cell Physiology, 2001, 280(2):C273-C279.
[26] Vafiadaki E, Arvanitis DA, Papalouka V, et al.Muscle lim protein isoform negatively regulates striated muscle actin dynamics and differentiation[J]. FEBS Journal, 2014, 281(14):3261-3279.

牦牛CSRP3基因的克隆及组织表达分析

Cloning and Tissue Expression Analysis of CSRP3 in Yak

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