牛源CCR7基因克隆、生物信息学及表达分析

doi:10.16431/j.cnki.1671-7236.2022.04.005

Abstract

Abstract: 【Objective】 This study was aimed to explore the biological characteristics of the protein encoded by CC chemokine receptor 7(CCR7) gene and the expression of mRNA in breast tissues of healthy and inflammatory dairy cows.【Method】 The CDS region of CCR7 gene in Chinese Holstein dairy cows was amplified by PCR method and cloned,the similarity alignment and phylogenetic tree construction were carried out after sequencing.The composition,physicochemical properties and structure of CCR7 protein were analyzed by bioinformatics softwares.The expression difference of CCR7 gene in breast tissues between healthy and inflammatory dairy cows were detected by Real-time quantitative PCR.【Result】 The full-length of CCR7 gene CDS region in Chinese Holstein dairy cows was 1 325 bp,which had high similarity (95.2%) and close genetic relationship with sheep;And had low similarity (56.4%) and far genetic relationship with chicken.CCR7 gene encoded 379 amino acids,among which the content of leucine was the most,the content of valine was the second,and the content of histidine and tryptophan were the lowest.The molecular weight of CCR7 protein was 14.56 ku,the theoretical isoelectric point was 8.89,and the average hydrophilic coefficient was 0.640,it was a transmembrane hydrophobic protein.The cleavage site of signal peptide might exist between amino acids 24 and 25.The structure of alpha helix was the most in the secondary structure,as high as 51.72%,and the tertiary structure was mainly alpha helix and random coil.The expression of CCR7 gene in breast tissue of inflammatory dairy cows was extremely significantly higher than that in healthy dairy cows (P<0.01),which might play an important role in the immune regulation of dairy cows mastitis.【Conclusion】 The CDS region of bovine CCR7 gene was successfully cloned,and the corresponding bioinformatics analysis and tissue quantitative expression were carried out,which provided materials for further study of the specific functions of CCR7 gene,and was of great significance to explore the regulatory function of CCR7 gene in dairy cow mastitis.

Key words: dairy cows; CCR7 gene; cloning; bioinformatics analysis; expression

CLC Number:

SHEN Xiang, CHEN Linghui, YAO Jingjie, WANG Sanhu, BAI Yueyu, ZHANG Xiaojian. Cloning,Bioinformatics and Expression Analysis of Bovine CCR7 Gene[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1244-1252.

References

[1] ZLOTNIK A,YOSHIE O.Chemokines:A new classification system and their role in immunity[J].Immunity,2000,12:121-127.
[2] HUGHES C E,NIBBS R J B.A guide to chemokines and their receptors[J].FEBS,2018,285:2944-2971.
[3] LIM HERMAN D.Systematic assessment of chemokine signaling at chemokine receptors CCR4,CCR7 and CCR10[J].International Journal of Molecular Sciences,2021,22(8):4232-4232.
[4] TAMAGNE M,PAKDAMAN S,BARTOLUCCI P,et al.Whole-blood CCR7 expression and chemoattraction in red blood cell alloimmunization[J].British Journal of Haematology,2021,194(2):477-481.
[5] COMERFORD I,HARATA-LEE Y,BUNTING M D,et al.A myriad of functions and complex regulation of the CCR7/CCL19/CCL21 chemokine axis in the adaptive immune system[J].Cytokine & Growth Factor Reviews,2013,24(3):269-283.
[6] SANCHO M,VIEIRA J M,CASALOU C,et al.Expression and function of the chemokine receptor CCR7 in thyroid carcinomas[J].Journal of Endocrinology,2006,191(1):229-238.
[7] SU M L,CHANG T M,CHIANG C H,et al.Inhibition of chemokine (C-C motif) receptor 7 sialylation suppresses CCL19-stimulated proliferation,invasion and anti-anoikis[J].PLoS One,2014,9(6):e98823.
[8] MO M,ZHOU M,WANG L,et al.CCL21/CCR7 enhances the proliferation,migration,and invasion of human bladder cancer T24 cells[J].PLoS One,2015,10(3):e0119506.
[9] BOYLE S T,INGMAN W V,POLTAVETS V,et al.The chemokine receptor CCR7 promotes mammary tumorigenesis through amplification of stem-like cells[J].Oncogene,2016,35(1):105-115.
[10] BOYLE S T,GIENIEC K A,GREGOR C E,et al.Interplay between CCR7 and Notch1 axes promotes stemness in MMTV-PyMT mammary cancer cells[J].Molecular Cancer,2017,16(1):19.
[11] CHEN Y,SHAO Z,JIANG E,et al.CCL21/CCR7 interaction promotes EMT and enhances the stemness of OSCC via a JAK2/STAT3 signaling pathway[J].Journal of Cellular Physiology,2020,235(9):5995-6009.
[12] BIRKENBACH M,JOSEFSEN K,YALAMANCHILIR,et al.Epstein-Barr virus-induced genes:First lymphocyte-specific G protein-coupled peptide receptors[J].Journal of Virology,1993,67(4):2209-2220.
[13] YOSHIDA R,NAGIRA M,KITAURA M,et al.Secondary lymphoid-tissue chemokine is a functional ligand for the CC chemokine receptor CCR7[J].Journal of Biological Chemistry,1998,273(12):7118-7122.
[14] 程玉洁,刘娟.趋化因子受体CCR7依赖的树突状细胞迁移的调控机制[J].中国免疫学杂志,2020,36(8):1004-1008. CHENG Y J,LIU J.Molecular regulation of CCR7-mediated dendritic cell migration[J].Chinese Journal of Immunology,2020,36(8):1004-1008.(in Chinese)
[15] 吴学磊,王艳华.CCL21/CCR7轴与乳腺癌转移及治疗的相关研究进展[J].生物技术通讯,2019,30(3):425-429. WU X L,WANG Y H.Progress on the correlation between CCL21/CCR7 axis and breast cancer metastasis and therapy[J].Letters in Biotechnology,2019,30(3):425-429.(in Chinese)
[16] 廖宇圣,张姮,李晖,等.CCR7在人肝癌组织中的表达及其与临床病理特征的关系[J].华中科技大学学报(医学版),2017,46(6):669-672. LIAO Y S,ZHANG H,LI H,et al.Expression of chemokine receptor CCR7 in human liver cancer tissues and its correlation with clinicopathological characteristics[J].Acta Medicinae Universitatis Scientiae et Technologiae Huazhong,2017,46(6):669-672.(in Chinese)
[17] HAUSER M A,LEGLER D F.Common and biased signaling pathways of the chemokine receptor CCR7 elicited by its ligands CCL19 and CCL21 in leukocytes[J].Journal of Leukocyte Biology,2016,99(6):869-882.
[18] MISHAN M A,AHMADIANKIA N,BAHRAMI A R.CXCR4 and CCR7:Two eligible targets in targeted cancer therapy[J].Cell Biology International,2016,40(9):955-967.
[19] CHIARA L D,CREAN J.Emerging transcriptional mechanisms in the regulation of epithelial to mesenchymal transition and cellular plasticity in the kidney[J].Journal of Clinical Medicine,2016,5(1):6.
[20] ZHANG Y Y,LIU Z B,YE X G,et al.Iodine regulates G2M progression induced by CCL21 CCR7 interaction in primary cultures of papillary thyroid cancer cells with RET PTC expression[J].Molecular Medicine Reports,2016,14(4):3941-3946.
[21] TANG F,CHEN J N,et al.Expression of CCL21 by EBV-associated gastric carcinoma cells protects CD8⁺CCR7⁺T lymphocytes from apoptosis via the mitochondria-mediated pathway[J].Pathology,2018,50(6):613-621.
[22] 赵云霞,姜丽娜,赵瑾.趋化因子受体 7 在乳腺癌组织中的表达及其与血管内皮生长因子-D 的关系[J].中国组织化学与细胞化学杂志,2018,27(5):440-445. ZHAO Y X,JIANG L N,ZHAO J.Expression of CCR7 in breast carcinoma and its relationship with vascular endothelial growth factor-D[J].Chinese Journal of Histochemistry and Cytochemistry,2018,27(5):440-445.(in Chinese)
[23] 李春磊,裴艳志,邹志田,等.非小细胞肺癌组织中趋化因子受体CCR7的表达及其临床意义[J].医学信息,2018,31(17):14-16. LI C L,PEI Y Z,ZOU Z T,et al.Expression of chemokine receptor CCR7 in non-small cell lung cancer and its clinical significance[J].Journal of Medical Information,2018,31(17):14-16.(in Chinese)
[24] SALEM A.CCR7 as a therapeutic target in cancer[J].Biochimica et Biophysica Acta.Reviews on Cancer,2021,1875(1):188499.
[25] 刘家齐,赵正晓,魏颖,等.芍药苷对哮喘模型小鼠气道炎症趋化因子及受体的干预作用[J].中国实验动物学报,2016,24(5):460-464. LIU J Q,ZHAO Z X,WEI Y,et al.Inhibitory effect of paeoniflorin on inflammatory chemokines and their receptor in a mouse model of asthma[J].Acta Laboratorium Animalis Scientia Sinica,2016,24(5):460-464.(in Chinese)
[26] 庞捷,余克强.活动期类风湿关节炎患者外周血树突状细胞CCR5和CCR7的检测及意义[J]. 南方医科大学学报,2010,30(9):2067-2069. PANG J,YU K Q.Detection and its clinical value of CCR5 and CCR7 in dendritic cells from patients with active rheumatoid arthritis[J].Journal of Southern Medical University,2010,30(9):2067-2069.(in Chinese)
[27] VAN R K.CCL21/CCR7 signaling in macrophages promotes joint inflammation and Th17-mediated osteoclast formation in rheumatoid arthritis[J].Cellular and Molecular Life Sciences,2020,77(7):1387-1399.
[28] KAWASHIMA D.Augmented expression of secondary lymphoid tissue chemokine and EBI1 ligand chemokine in Crohn’s disease[J].Journal of Clinical Pathology,2005,58(10):1057-1063.

Cloning,Bioinformatics and Expression Analysis of Bovine CCR7 Gene

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