草原红牛肝配蛋白A5基因克隆及其在不同组织中的表达研究

doi:10.16431/j.cnki.1671-7236.2021.06.009

Abstract

Abstract: The purpose of this study was to explore the gene function of ephrin A5(EFNA5) in Chinese Red steppe,and to detect the difference of EFNA5 gene expression in different tissues of Chinese Red steppe by bioinformatics analyzed.Primers were designed according to the predicted sequence of EFNA5 gene in GenBank (accession No.:NM_001076432.1).Verification of complete coding sequence of EFNA5 gene was obtained by PCR and sequencing.Using analysis software to carry out sequence similarity alignment and construct phylogenetic tree.The corresponding amino acid sequence was obtained.The physical and chemical property and subcellular structure was analysed by online prediction software.Protein hydrophilicity,hydrophobicity and phosphorylation sites were analysed.Protein secondary structure was predicted and protein tertiary structure model was constructed.EFNA5 gene expression in tissues of Chinese Red steppe was detected by Real-time quantitative PCR.The results showed that the EFNA5 gene of Chinese Red steppe had the highest similartiy with common cattle and American bison (100% and 99.8%),and with sheep and human were 95.6% and 97.2% respectively,and lower similartiy with dolphins,chickens,horses,rhesus monkeys,mice and pigs (83.6%,85.8%,84.5%,84.9%,84.1% and 82.8%,respectively).The CDS of EFNA5 gene was 687 bp,encoding 228 amino acids.The molecular formula of the protein was C₁₁₈₃H₁₇₉₁N₃₁₅O₃₃₉S₁₃,the molecular weight was 26.266 ku,the theoretical isoelectric point was 5.97.The protein instability index was 49.66,and the total average hydrophilicity was －0.313. Phosphorylation sites prediction analysis showed that there were 27 phosphorylation sites in EFNA5 protein.The results of subcellular localization showed that EFNA5 protein was mainly located in cytoplasm (26.1%),secretory system vesicles (27.1%),nucleus (13.0%),mitochondria (13.0%),plasma membrane (8.7%),endoplasmic reticulum (4.3%),golgi apparatus (4.3%),cytoskeleton (4.3%) and vacuole (4.3%).The main forms of secondary structure were α-helix (18.9%),β-turn (30.2%),β-fold (26.4%) and irregular crimp (24.5%),which were consistent with the prediction of tertiary structure.The expression of EFNA5 gene was the highest in kidney and stomach,and the least in lung.There was significant difference in different tissues of Chinese Red steppe (P<0.05).This study could provide reference for further investigation of the effects of EFNA5 gene on lipid metabolism in livestock and the screening of quality candidate genes in Red steppe.

Key words: Chinese Red steppe; EFNA5 gene; cloning; bioinformatics analysis; expression difference

CLC Number:

XUE Jiajia, QIN Lihong, CAO Yang, GAO Yi, LIU Lixiang, LI Xu, XIAO Cheng, WANG Jing, ZHANG Guoliang. Cloning of Ephrin A5 Gene and Its Expressing in Different Tissues in Red Steppe[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 1967-1975.

References

[1] 胡成华,张国梁,吴健,等.草原红牛泌乳与产肉性能选育研究[J].现代农业科技,2009,5:210-211. HU C H,ZHANG G L,WU J,et al.Study on breeding of lactation and meat production performance of Red steppe[J].Modern Agricultural Science and Technology,2009,5:210-211.(in Chinese)
[2] 张国梁,李旭,吴健,等.关于中国草原红牛发展的思考[J].吉林畜牧兽医,2013,34(11):12-13. ZHANG G L,LI X,WU J,et al.Thoughts on the development of Red steppe[J].Jilin Animal Husbandry and Veterinary Medicine,2013,34 (11):12-13.(in Chinese)
[3] WANG T H,YEH C T,HO J Y,et al.OncomiR miR-96 and miR-182 promote cell proliferation and invasion through targeting ephrinA5 in hepatocellular carcinoma[J].Molecular Carcinogenesis,2016,55(4):366-375.
[4] WANG T H,CHANG J L,HO J Y,et al.EphrinA5 suppresses colon cancer development by negatively regulating epidermal growth factor receptor stability[J].The FEBS Journal,2012,279(2):251-263.
[5] LI J J,LIU D P,LIU G T,et al.EphrinA5 acts as a tumor suppressor in glioma by negative regulation of epidermal growth factor receptor[J].Oncogene,2009,28(15):1759-1768.
[6] WORKU T,WANG K,AYERS D,et al.Regulatory roles of ephrinA5 and its novel signaling pathway in mouse primary granulosa cell apoptosis and proliferation[J].Cell Cycle,2018,17(7):892-902.
[7] LIN S,LIU W,CHEN C L,et al.Neogenin-loss in neural crest cells results in persistent hyperplastic primary vitreous formation[J].Molecular Biology of the Cell,2020,12(1):17-31.
[8] SZEPIETOWSKA B,HORVATH T L,SHERWIN R S.Role of synaptic plasticity and ephA5-ephrinA5 interaction within the ventromedial hypothalamus in response to recurrent hypoglycemia[J].Diabetes,2014,63(3):1140-1147.
[9] 徐露.产热脂肪细胞(Aldhlal^-/-细胞)分泌物对白色脂肪神经支配及轴突生长的调控作用[D].苏州:苏州大学,2016. XU L.Secretome from thermogenic adipocytes(Aldhlal^-/- cells) regulates axonal growth and innervation of white adipose tissue[D].Suzhou:Suzhou University,2016.(in Chinese)
[10] AN B,XU L,XIA J,et al.Multiple association analyzed of loci and candidate genes that regulate body size at three growth stages in Simmental beef cattle[J].BMC Genetics,2020,21(1):21-32.
[11] BOLZ J,UZIEL D,MÜHLFRIEDEL S,GÜLLMAR A,et al.Multiple roles of ephrins during the formation of thalamocortical projections:Maps and more[J].Journal of Neurobiology,2004,59(1):82-94.
[12] OTAL R,BURGAYA F,FRISÉN J,et al.Ephrin-A5 modulates the topographic mapping and connectivity of commissural axons in murine hippocampus[J].Neuroscience,2006,141(1):109-121.
[13] HERATH N I,SPANEVELLO M D,SABESAN S,et al.Over-expression of Eph and ephrin genes in advanced ovarian cancer:Ephrin gene expression correlates with shortened survival[J].BMC Cancer,2006,6:144.
[14] VARELIAS A,KOBLAR S A,COWLED P A,et al.Human osteosarcoma expresses specific ephrin profiles:Implications for tumorigenicity and prognosis[J].Cancer,2002,95(4):862-869.
[15] RICCI B,MILLNER T O,POMELLA N,et al.Polycomb-mediated repression of ephrinA5 promotes growth and invasion of glioblastoma[J].Oncogene,2020,39(12):2523-2538.
[16] JIANG N,DAI Q,SU X,et al.Role of PI3K/Akt pathway in cancer:The framework of malignant behavior[J].Molecular Biology Reports,2020,47(6):4587-4629.
[17] YUN J,JIN H,CAO Y,et al.RNA-seq analyzed reveals a positive role of HTR2A in adipogenesis in Yan Yellow cattle[J].International Journal of Molecular Sciences,2018,19(6):1760.
[18] LIU R R,SUN Y F,ZHAO G P,et al.Identification of loci and genes for growth related traits from a genome-wide association study in a slow-x fast-growing broiler chicken cross[J].Genes and Genomics,2015,37(10):829-836.
[19] NIKOLOV D,LI C,LACKMANN M,et al.Crystal structure of the human ephrin-A5 ectodomain[J].Protein Science,2007,16(5):996-1000.
[20] DAVY A,GALE N W,MURRAY E W,et al.Compartmentalized signaling by GPI-anchored ephrin-A5 requires the Fyn tyrosine kinase to regulate cellular adhesion[J].Genes & Development,1999,13(23):3125-3135.
[21] WIJERATNE D,RODGER J,STEVENSON A,et al.Ephrin-A2 affects wound healing and scarring in a murine model of excisional injury[J].Burns,2019,45(3):682-690.
[22] PASQUALE E B.Eph-ephrin bidirectional signaling in physiology and disease[J].Cell,2008,133(1):38-52.
[23] LIN L,KIGHTLINGER W,PRABHU S K,et al.Sequential glycosylation of proteins with substrate-specific N-glycosyltransferases[J].ACS Central Science,2020,6(2):144-154.
[24] HAFNER C,SCHMITZ G,MEYER S,et al.Differential gene expression of Eph receptors and ephrins in benign human tissues and cancers[J].Clinical Chemistry,2004,50(3):490-499.
[25] ISLAM S,LOIZIDES A M,FIALKOVICH J J,et al.Developmental expression of Eph and ephrin family genes in mammalian small intestine[J].Digestive Diseases and Sciences,2010,55(9):2478-2488.

Cloning of Ephrin A5 Gene and Its Expressing in Different Tissues in Red Steppe

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LIU Jialong, ZHANG Yiwen, ZHANG Yueran, SUN Qijuan, CHEN Jian, HE Lei, JIA Yanyan, DING Ke, YU Zuhua. Cloning,Prokaryotic Expression and Bioinformatics Analysis of ELAVL1 Gene in Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1807-1817.
[2]	ZHUO Na, ZHU Zhongwu, LI Jing, HU Shixiang, WANG Jianchang, LIN Hua, CHEN Chaolin, HAN Diangang, AI Jun, CHEN Peifu. Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1959-1970.
[3]	FAN Bingfeng, LI Wen, LIU Lixiang, ZHAO Xiangyuan, SHAO Jing, LIN Lefeng, SUN Huimin, ZHANG Xulin, XU Baozeng. Cloning and Bioinformatics Analysis of AANAT Gene in Mink [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1307-1318.
[4]	LIU Hongrun, ZHU Siran, FENG Lingli, ZHANG Kun, YAN Gang, WANG Yubin, ZHANG Shuai, JIANG Shan, XU Di, LAN Ganqiu, LIANG Jing. Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1352-1363.
[5]	JIA Xinyue, MA Jing, ZHANG Yanyan, MA Xun, WANG Zhengrong, BO Xinwen. Bioinformatics Analysis and Prokaryotic Expression of EGR-07243 and EGR-07243 Genes of Echinococcus granulosus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 847-858.
[6]	ZHANG Yongjie, SUI Zhiyuan, ZHANG Zhishuai, WANG Chenguang, LI Xiaojun, XING Feng. Cloning and Bioinformatics Analysis of NPTX1 Gene and Its Tissues Expression at Puberty in Dolang Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1048-1058.
[7]	WEI Yirong, ZHENG Zihua, ZHANG Sanbao, SONG Ying, JIANG Haiyu, SUN Wenyue, CHENG Pengfei, LIU Yufan, ZOU Jianwei, HUANG Yanna, PAN Yan, JIANG Qinyang. Cloning,Bioinformatics Analysis and Tissue Expression of UCP1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 440-450.
[8]	TANG Jingyu, DU Hanyu, MENG Chunchun, LIU Guangqing. Cloning,Bioinformatics Analysis and Subcellular Localization of Interferon-stimulated Gene 15 in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2843-2854.
[9]	FU Yongzhi, MA Xiaojun, WANG Maohan, LI Zhe, CHEN Zhiyuan, QUAN Wei, ZHAO Han, SU Yongmei, WU Jianhua, ZHU Wenjin. Bioinformatics Analysis and Tissue Expression of MSTN Gene in Yangyuan Donkeys [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2910-2919.
[10]	QI Nannan, QIAN Yonghang, LUO Gang, SITU Xuqi, LIU Jiying. Cloning, Bioinformatics Analysis and Tissue Expression Location of Porcine MORC2 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2431-2441.
[11]	ZHANG Hang, SHA Huiyang, LI Huawei, HUANG Liangzong, ZHAO Mengmeng. Cloning, Bioinformatics and Tissue Expression Analysis of TRIM3 Gene in Swine [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2474-2483.
[12]	ZOU Jianwei, ZOU Juhong, SHEN Yujian, WEI Yi, ZHANG Sanbao, LIAN Zitong, XU Jianjian, SONG Ying, HUANG Yanna, WEI Yingming, JIANG Qinyang, ZHENG Zihua. Bioinformatics Analysis, Construction of Eukaryotic Expression Vector and Expression of LMCD1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2033-2042.
[13]	YAN Yelian, ZHANG Mengya, LIU Qiuchen, WANG Xin, XU Changzhi, ZONG Yanfeng, ZHU Zhihua, WU Sucheng, SONG Yu, LI Yunsheng, ZHANG Yunhai, CAO Zubing. Research Progress of Epigenetic Reprogramming in Mammalian Somatic Cell Nuclear Transfer Embryo Development [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2259-2269.
[14]	ZHANG Limeng, LIU Aiju, LI Runting, LI Yuhua, LI Lin, NIE Xiaoning, WANG Linqing, CHEN Longxin. Cloning,Bioinformatics and Tissue Expression Analysis of CREBRF Gene in Ovis aries [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1599-1609.
[15]	WANG Baobao, JIN Hang, XIAN Yuhan, FENG Hongsheng, GAO Fengshan. Cloning SLA-1 Gene from Porcine Kidney Epithelial Cells and Analyzing its Molecular Structure Characteristics [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1671-1678.