合作猪CXCL12基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2025.06.003

Abstract

Abstract: 【Objective】 This study was aimed to clone and perform bioinformatics analysis on the stromal cell-derived factor-1 (SDF-1/CXCL12) gene in Hezuo pigs，and detect the expression of CXCL12 gene in different tissues,establishing a foundation for further functional investigation of CXCL12 gene.【Method】 Using cDNA from the heart of Hezuo pigs as a template,the CXCL12 gene CDS sequence was amplified via PCR,cloned and sequenced,the similarity alignment and phylogenetic tree construction for the obtained sequence were performed.Bioinformatics software was used to analyze the physicochemical property and structural function of CXCL12 protein.The expression of CXCL12 gene in different tissues of Hezuo pigs was detected by Real-time quantitative PCR.【Result】 The full-length CDS sequence of CXCL12 gene in Hezuo pigs was 351 bp,encoding 116 amino acids.CXCL12 gene in Hezuo pigs was most closely related to Equus cabllus,and was distantly related to Oryx dammah.CXCL12 protein lacked transmembrane regions and signal peptide,included a low complexity region SCY,with a hydrophilicity index of ―0.347,it belonged to hydrophilic proteins.CXCL12 protein contained 12 phosphorylation sites.The secondary structure was predominantly compose of random coil (54.31%) and alpha helix (32.76%),which was consistent with the tertiary structure.The protein was primarily located in nucleus and mitochondria,and interacted with proteins such as CCL21 and CXCL9.Real-time quantitative PCR results showed that the expression of CXCL12 gene in liver of Hezuo pigs was extremely significantly higher than that in other tissues (P＜0.01),and the expression in kidney and heart was extremely significantly lower than that in other tissues (P＜0.01).【Conclusion】 This study successfully cloned the CDS sequence of CXCL12 gene in Hezuo pigs,encoding 116 amino acids,mainly distributed in nucleus and mitochondria.It was a stable,hydrophilic and secretory protein without transmembrane structures.The highest expression of CXCL12 gene was observed in liver of Hezuo pigs.The results provided a reference for further exploring the regulatory roles of CXCL12 gene in immune mechanisms of Hezuo pigs.

Key words: Hezuo pigs; CXCL12 gene; cloning; bioinformatics; expression

CLC Number:

S813.3
Q78

YANG Quan, LI Xiao, YAN Zunqiang, WANG Pengfei, HUANG Xiaoyu, GAO Xiaoli, YANG Qiaoli, GUN Shuangbao, YANG Jiaojiao. Cloning,Bioinformatics Analysis and Tissue Expression of CXCL12 Gene in Hezuo Pigs[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2482-2493.

References

[1] JANSSENS R,STRUYF S,PROOST P.The unique structural and functional features of CXCL12[J].Cellular & Molecular Immunology,2018,15(4):299-311.
[2] ZHOU W,GUO S,LIU M.et al.Targeting CXCL12/CXCR4 axis in tumor immunotherapy[J].Current Medicinal Chemistry,2019,26(17):3026-3041.
[3] MULLER A,HOMEY B,SOTO H.et al.Involvement of chemokine receptors in breast cancer metastasis[J].Nature,2001,410(6824):50-56.
[4] CAMBIER S,GOUWY M,PROOST P.The chemokines CXCL8 and CXCL12:Molecular and functional properties,role in disease and efforts towards pharmacological intervention[J].Cellular & Molecular Immunology, 2023,20(3):217-251.
[5] 张敏.趋化因子CXCL12对牛和猪卵母细胞成熟及胚胎发育影响的研究[D].杨凌：西北农林科技大学,2023.ZHANG M.Effects of chemokine CXCL12 on oocyte maturation and embryo development in bovine and porcine[D].Yangling:Northwest A&F University,2023.(in Chinese)
[6] 宫玉霞,姚铭,李红梅，等.合作猪资源保护利用现状及发展建议[J].国外畜牧学(猪与禽),2019,39(5):35-37.GONG Y X,YAO M,LI H M,et al.Current situation and development suggestions of conservation and utilization of Hezuo pigs resources[J].Animal Science Abroad-Pigs and Poultry,2019,39(5):35-37.(in Chinese)
[7] 马莉.浅谈甘肃迭部县蕨麻猪养殖优势[J].中国猪业,2023,18(4):35-37.MA L.A brief discussion on the breeding advantages of Juema pigs in Diebu county,Gansu province[J].China Swine Industry,2023,18(4):35-37.(in Chinese)
[8] 陈国顺,滚双宝.合作猪种质资源调查研究[J].猪业科学,2013,30(9):124-127.CHEN G S,GUN S B.Investigation and study on Hezuo pig germplasm resources[J].Swine Industry Science,2013,30(9):124-127.(in Chinese)
[9] 周琳,王蜀金.藏猪生理学研究进展[J].家畜生态学报,2014,35(7):7-12.ZHOU L,WANG S J.Research progress on Tibetan swine physiology[J].Acta Ecologae Animalis Domastici,2014,35(7):7-12.(in Chinese)
[10] 杨延彪,曹国顺,郭宏伟,等.甘南蕨麻猪的生态分布及其生产性能[J].家畜生态学报,2003,24(1):75-76.YANG Y B,CAO G S,GUO H W,et al.Ecological distribution and productive performance of Juema pig in Southern part of Gansu province[J].Acta Ecologae Animalis Domastici, 2003,24(1):75-76.(in Chinese)
[11] 李富强,马小军,张小丽,等.合作猪生理生化及免疫指标研究[J].甘肃农业大学学报,2013,48(3):24-27.LI F Q,MA X J,ZHANG X L,et al.Characteristics of blood physiological and biochemical and immunological indexes of Hezuo swine[J].Journal of Gansu Agricultural University,2013,48(3):24-27.(in Chinese)
[12] 刘燕,刘孟洲.合作猪血液生理生化指标特性的研究[J].中国畜牧兽医,2007,34(7):52-55.LIU Y,LIU M Z.The study on haemal physiological and biological parameters of Hezuo swine [J].China Animal Husbandry & Veterinary Medicine,2007,34(7):52-55.(in Chinese)
[13] 张爱文,文璟,李康伟.甘肃省生猪种业现状及发展策略初探[J].中国畜禽种业,2023,19(11):21-25.ZHANG A W,WEN J,LI K W.Preliminary study on the current situation and development strategy of pig seed industry in Gansu province[J]. The Chinese Livestock and Poultry Breeding,2023,19(11):21-25.(in Chinese)
[14] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using Real-time quantitative PCR and the 2(-delta delta C(t)) method[J]. Methods (San Diego,Calif),2001,25(4):402-408.
[15] MOUSAVI A.CXCL12/CXCR4 signal transduction in diseases and its molecular approaches in targeted-therapy[J].Immunology Letters,2020,217:91-115.
[16] 庹锐.黄鳝免疫基因CXCR4和IL-1β及其受体的克隆和表达研究[D].荆州：长江大学,2014.TUO R.Cloning and expression of immune genes CXCR4 and IL-1β and their receptors in Monopterus albus[D].Jingzhou:Yangtze University,2014.(in Chinese)
[17] 蒋玉萍,吴小华.趋化因子CXCL12及其受体与肿瘤转移的关系[J].癌症,2007,2:220-224.JIANG Y P,WU X H.Correlations of chemokine CXCL12 and its receptor to tumor metastasis[J].Journal of Cancer,2007,2:220-224.(in Chinese)
[18] AGERER B,LERCHER A,BERGTHALER A.The serine's call:Suppressing interferon responses[J].Cell Metabolism, 2021,33(5):849-850.
[19] GHAYOURI M,BOULWARE D,NASIR A.et al.Activation of the serine/theronine protein kinase Akt in enteropancreatic neuroendocrine tumors[J].Anticancer Research,2010,30(12):5063-5068.
[20] MAKOWSKA M,WARDOWSKA A,BAUER M,et al.Impact of lipidation site on the activity of α-helical antimicrobial peptides[J].Bioorganic Chemistry,2024,153:107821.
[21] NGUYEN T,LAGMEN C,CHUNG L K.et al.Insights into CCL21's roles in immunosurveillance and immunotherapy for gliomas[J]. Journal of Neuroimmunology,2017,305:29-34.
[22] SALEHI-RAD R,LIM R J,DU Y.et al.CCL21-DC in situ vaccination in murine NSCLC overcomes resistance to immunotherapy and generates systemic tumor-specific immunity[J].Journal for Immunotherapy of Cancer,2023,11(9):e006896.
[23] NEO S Y,LUNDQVIST A.The multifaceted roles of CXCL9 within the tumor microenvironment[J].Advances in Experimental Medicine and Biology,2020,1231:45-51.
[24] QUINN K E,MACKIE D I,CARON K M.Emerging roles of atypical chemokine receptor 3(ACKR3) in normal development and physiology[J]. Cytokine,2018,109:17-23.
[25] ANASTASIADOU D P,COUTURIER N,GOEL S,et al.Intratumoral CXCL12 gradients contextualize tumor cell invasion,migration and immune suppression in breast cancer[J].BioRxiv:The Preprint Server for Biology,2024，18:618571.
[26] PESSENDA G,FERREIRA R T,PAUN A,et al.Kupffer cell and recruited macrophage heterogeneity orchestrate granuloma maturation and hepatic immunity in visceral leishmaniasis.[J].BioRxiv： The Preprint Server for Biology, 2024,13:602717.
[27] HOMAN A E,GILANI A,NAVARRO R A,et al.Complement 3a receptor 1 on macrophages and kupffer cells is not required for the pathogenesis of metabolic dysfunction-associated steatotic liver disease[J].MedRxiv:The Preprint Server for Health Sciences,2024,9:24309550.
[28] 陈宏远,谭毅,马伟峰,等.CXCL12-α基因编码区的克隆与生物信息学分析[J].生物技术,2014,24(3):9-16.CHEN H Y,TAN Y,MA W F,et al.Cloning and bioinformatics analysis of human CXCL12-α gene coding region[J].Biotechnology,2014,24(3):9-16.(in Chinese)
[29] 莫斯科,房永祥,冯海燕,等.甘肃合作猪白细胞抗原经典Ⅱ类分子DR基因克隆及生物信息学分析[J].畜牧兽医学报,2009,40(11):1662-1668.MO S K,FANG Y X,FENG H Y,et al.Molecular cloning and bioinformatics analysis of swine leukocyte antigen class Ⅱ DR genes in Gansu Hezuo pigs[J]. Acta Eterinaria et Zootechnica Sinica,2009,40(11):1662-1668.(in Chinese)
[30] MEHRPOURI M.The contributory roles of the CXCL12/CXCR4/CXCR7 axis in normal and malignant hematopoiesis:A possible therapeutic target in hematologic malignancies[J].European Journal of Pharmacology,2022,920:174831.
[31] TIMOTIJEVIC G,MOSTARICA STOJKOVIC M,MILJKOVIC D.CXCL12:Role in neuroinflammation[J].International Journal of Biochemistry and Cell Biology,2012,44(6):838-841.
[32] 李艳,杨义然,孙英,等.趋化因子CXCL12对2型固有淋巴细胞定向趋化的作用研究[J].中国耳鼻咽喉头颈外科,2019,26(11):609-612.LI Y,YANG Y R,SUN Y,et al.Potential role of chemokine CXCL12 on chemotaxis of group 2 innate lymphoid cells[J].Chinese Archives of Otolaryngology-Head and Neck Surgery,2019,26(11):609-612.(in Chinese)
[33] MA J,SU H,YU B,et al.CXCL12 gene silencing down-regulates metastatic potential via blockage of MAPK/PI3K/AP-1 signaling pathway in colon cancer[J].Clinical & Translational Oncology,2018,20(8):1035-1045.
[34] PING Y F,YAO X H,JIANG J Y,et al.The chemokine CXCL12 and its receptor CXCR4 promote glioma stem cell-mediated VEGF production and tumour angiogenesis via PI3K/Akt signalling[J].Journal of Pathology,2011,224(3):344-354.
[35] YAMADA K,MAISHI N,AKIYANA K et al.CXCL12-CXCR7 axis is important for tumor endothelial cell angiogenic property[J].International Journal of Cancer,2015,137(12):2825-2836.
[36] YANG Y,LI J,LEI W.et al.CXCL12-CXCR4/CXCR7 axis in cancer:From mechanisms to clinical applications[J].International Journal of Biological Sciences,2023,19(11):3341-3359.
[37] 庞同涛.骨质疏松发病与Bcl-2、CXCL12基因表达的关系[D].济南：山东大学,2019.PANG T T.Relationship between osteoporosis and expressions of Bcl-2 and CXCL12[D].Jinan:Shandong University,2019.(in Chinese)

Cloning,Bioinformatics Analysis and Tissue Expression of CXCL12 Gene in Hezuo Pigs

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	GUAN Song, SHI Liguang, LIN Yu, JIANG Jianxiao, WU Hongzhi, PENG Weiqi. Functional Prediction of GDF9 Gene nsSNP and Its Correlation Analysis with Litter Size in Hainan Black Goats [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2166-2176.
[2]	LI Tianxiu, LI Xinpeng, DONG Xinxing, LAN Guoxiang, YAN Dawei, ZHU Jiawei. Amplification,Sequence Analysis and Tissue Expression Study of HMOX2 Gene in Lijiang Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2219-2231.
[3]	LIU Min, CHENG Huai, ZHENG Zhenzhen, ZHANG Mengsi, ZHANG Hewei, REN Jingqiang. Eukaryotic Expression and Polyclonal Antibody Preparation of N Protein of Canine Parainfluenza Virus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2287-2294.
[4]	HU Huihui, FU Panpan, LI Jie, YAN Zunqiang, GAO Xiaoli, YANG Jiaojiao, HUANG Xiaoyu. Cloning,Identification and Tissue Expression Analysis of TRIF Gene CDS Region in Hezuo Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1478-1487.
[5]	LIU Xiaotong, GE Bingjie, QIU Qian, LIU Xinman, YU Minghong, ZHANG Xuemei. Application of Bioinformatics in Toxicity Mechanism of Aflatoxin B₁ [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1639-1650.
[6]	ZHANG Ying, CHEN Guangzhen, LIU Mei, LI Hongxia, YU Yan, JIA Xuebo, LI Wenqian, FAN Jianhua, MA Zhiyu. Cloning,Bioinformatics and Tissue Expression Analysis of NMB Gene in Mulard Duck [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1705-1715.
[7]	SI Wei, TAN Xiyu, XU Lingyun, LUO Tingrong, LI Xiaoning, GU Jinyan. Prokaryotic Expression of HA1 Protein of Swine Influenza Virus Subtype H3N2 and Preparation of Polyclonal Antibodies [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1739-1749.
[8]	MIAO Shukui, MI Xiaoyun, WEI Jie, WEI Yurong, HAILIQIEMU·Maimaitiyiming. Cloning and Bioinformatics Analysis of VP7 Gene of the Genotype G8 Bovine Rotavirus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1784-1795.
[9]	HUANG Nianni, HUANG Zike, LI Congcong, LIU Han, MA Shiyu, SONG Guo, JIANG Liang, PENG Weibo, YANG Lianxi, HUANG Shengqiang. Cloning,Bioinformatics and Tissue Expression Analysis of FGF10 Gene in Jiuyishan Rabbits [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(3): 1001-1010.
[10]	WANG Yong, NA Risu, LIANG Jianyong, LIU Yang, LI Jiuyue, GAO Yuan, TIAN Feng, XUE Shuyuan. Effects of Quercetin on Follicular Development,Follicular Fluid Microenvironment and Ovarian Steroidogenesis Related Gene Expression in Mongolian Sheep [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(3): 1150-1158.
[11]	PAN Xiangying, ZENG Zhiyong, LIANG Haiying, TANG Deyuan, WANG Bin, YE Ni, TIAN Hongli, BIAN Mengting, LIU Jiajia, HUANG Shu. Truncated Expression of Porcine Rotavirus VP6 Protein and Establishment of an Indirect ELISA Antibody Detection Method [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(3): 1231-1240.
[12]	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 & Veterinary Medicine, 2025, 52(2): 499-511.
[13]	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 & Veterinary Medicine, 2025, 52(2): 534-544.
[14]	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 & Veterinary Medicine, 2025, 52(2): 740-748.
[15]	ZHAO Wenqing, BO Xinwen, PU Na, ZHANG Yuxia, CHEN Xuke, ZHANG Yanyan, SUN Yan, QI Meng, WANG Zhengrong. Bioinformatics Analysis and Prokaryotic Expression of EgBAL Protein of Echinococcus granulosus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(2): 801-810.