OPN基因序列分析及其在产蛋后期鸡胫骨中的表达研究

doi:10.16431/j.cnki.1671-7236.2019.08.025

Abstract

Abstract:

To investigate the structure and function of osteopontin (OPN) gene and explore its expression characteristics in bone metabolism of layer during late period of laying.The tibia of Hyline Grey layer at late period of laying was selected for RNA isolation.The specific primers were designed based on OPN gene sequence of Gallus gallus in GenBank (accession No.:NC_006091.5) using Primer Premier 3.0 online software.The coding sequence (CDS) of OPN gene was cloned by RT-PCR and analyzed by bioinformatics.Real-time PCR technology was used to detect the expression of OPN gene mRNA in tibia.The results showed that the full-length of chicken OPN gene CDS was 795 bp, encoding 264 amino acids.The phylogenetic tree constructed by Mega 6.0 software showed that the relationship was the closest between chicken and carp, with 100% similarity.The OPN gene was highly conserved in avian evolution.The bioinformatics analysis suggested that OPN was an unstable and hydrophilic protein with 27 O-glycosylation sites, 1 N-glycosylation site and 34 phosphorylation sites.The subcellular localization of OPN was mainly localized in cytoplasm (44.4%) and mitochondria (33.3%).The secondary structure was mainly composed of random coil and alpha helix.The difference of OPN gene mRNA expression among individuals was not related to bone strength.The results indicated that OPN gene mRNA expression was not related to the bone strength, which provided a reference basis for further study on the mechanism of OPN gene in bone metabolism of layer during late period of laying.

Key words: layer; tibia; OPN gene; late period of laying

CLC Number:

S813.3

CHEN Ye, HOU Jianku, HOU Fei, LI Xianglong, XU Lijun, WANG Han, CHEN Hui, ZHOU Rongyan. Sequence and Expression Analysis of OPN Gene in Tibia of Layer During Late Period of Laying[J]. , 2019, 46(8): 2397-2405.

References 28

[1]	黄淑纾,林华,朱秀芬,等.骨质量与骨质疏松性骨折[J].中华骨质疏松和骨矿盐疾病杂志,2012,5(4):285-291. HUANG S Y,LIN H,ZHU X F,et al.Bone quality and osteoporotic fracture[J]. Chinese Journal of Osteoporosis and Bone Mineral Research,2012,5(4):285-291.(in Chinese)
[2]	周振雷,胡云峰,陈秀霞,等.笼养蛋鸡骨质疏松症研究进展[J].畜牧与兽医,2007,39(8):71-74. ZHOU Z L,HU Y F,CHEN X X,et al.Research progress on osteoporosis in caged laying hens[J]. Animal Husbandry & Veterinary Medicine,2007,39(8):71-74.(in Chinese)
[3]	马利芹,利凯,吕文亭,等.植物雌激素对笼养蛋鸡疲劳综合症骨骼重塑作用机制的研究进展[J].养禽与禽病防治,2009,10:2-4. MA L Q,LI K,LYU W T,et al.Advances in research on the mechanism of phytoestrogens on bone remodeling in caged hens fatigue syndrome[J]. Poultry Husbandry and Disease Control,2009,10:2-4.(in Chinese)
[4]	马伟光,周树夏,刘宝林.PTH对破骨细胞骨吸收功能的影响及成骨细胞介导作用[J].中国病理生理杂志,1995,1:102-105. MA W G,ZHOU S X,LIU B L.Effect of PTH on bone resorption of osteoclasts and osteoblast-mediated[J]. Chinese Journal of Pathophysiology,1995,1:102-105.(in Chinese)
[5]	主性,沈向真,侯加法,等.血浆BGP、AKP对笼养蛋鸡骨质疏松症的诊断学意义[J].中国家禽,2000,5:6-9. ZHU X,SHEN X Z,HOU J F,et al.Study on significance of plasma BGP,AKP,calcium and phosphorusfor diagnosis of cage layer osteoporosis[J]. China Poultry,2000,5:6-9.(in Chinese)
[6]	WHITEHEAD C C.Overview of bone biology in the egg-laying hen[J]. Poultry Science,2004,83(2):193-199.
[7]	HERNANDEZ C J,KEAVENY T M.A biomechanical perspective on bone quality[J]. Bone,2006,39(6):1173-1181.
[8]	ZIOUPOS P,CURREY J D,HAMER A J.The role of collagen in the declining mechanical properties of aging human cortical bone[J]. Journal of Biomedical Materials Research,1999,45(2):108-116.
[9]	FRATZL P,GUPTA H S,PASCHALIS E P,et al.Structure and mechanical quality of the collagen-mineral nano-composite in bone[J]. Journal of Materials Chemistry,2004,14(14):2115-2123.
[10]	ROSCHGER P,PASCHALIS E P,FRATZL P,et al.Bone mineralization density distribution in health and disease[J]. Bone,2008,42(3):456-466.
[11]	BUSSE B,HAHN M,SOLTAU M,et al.Increased calcium content and inhomogeneity of mineralization render bone toughness in osteoporosis:Mineralization,morphology and biomechanics of human single trabeculae[J]. Bone,2009,45(6):1034-1043.
[12]	SILVA M J,BRODT M D,WOPENKA B,et al.Decreased collagen organization and content are associated with reduced strength of demineralized and intact bone in the SAMP6 mouse[J]. Journal of Bone and Mineral Research,2006,21(1):78-88.
[13]	TANG S Y,ZEENATH U,VASHISHTH D.Effects of non-enzymatic glycation on cancellous bone fragility[J]. Bone,2007,40(4):1144-1151.
[14]	FANTNER G E,HASSENKAM T,KINDT J H,et al.Sacrificial bonds and hidden length dissipate energy as mineralized fibrils separate during bone fracture[J]. Nature Materials,2005,4(8):612-616.
[15]	HUANG W,CARLSEN B,RUDKIN G,et al.Osteopontin is a negative regulator of proliferation and differentiation in MC3T3-E1 pre-osteoblastic cells[J]. Bone,2004,34(5):799-808.
[16]	茅文斌,邵增务.骨桥蛋白与骨质疏松症研究进展[J].中国骨质疏松杂志,2007,13(3):204-207. MAO W B,SHAO Z W.Research development of osteopontin and osteoporosis[J]. Chinese Journal of Osteoporosis,2007,13(3):204-207.(in Chinese)
[17]	SHEVDE L A,SAMANT R S.Role of osteopontin in the pathophysiology of cancer[J]. Matrix Biology,2014,37(37):131-141.
[18]	邹呈雨,宋关斌.骨桥蛋白促细胞迁移作用及其分子机理[J].生命的化学,2009,29(4):543-546. ZOU C Y,SONG G B.Osteopontin promotes cell migration and its molecular mechanism[J]. Chemistry of Life,2009,29(4):543-546.(in Chinese)
[19]	DENHARDT D T,NODA M.Osteopontin expression and function:Role in bone remodeling[J]. Journal of Cellular Biochemistry,2015,72(30):92-102.
[20]	NAU G J,CHUPP G L,EMILE J F,et al.Osteopontin expression correlates with clinical outcome in patients with mycobacterial infection[J]. American Journal of Pathology,2000,157(1):37-42.
[21]	KANG K A,PIAO M J,RYU Y S,et al.Cytoplasmic localization of RUNX3 via histone deacetylase-mediated SRC expression in oxidative-stressed colon cancer cells[J]. Journal of Cellular Physiology,2017,232(7):1914-1921.
[22]	GREGORY N G,WILKINS L J.Broken bones in domestic fowl:Handling and processing damage in end-of-lay battery hens[J]. British Poultry Science,1989,30(3):555-562.
[23]	WAN Y M,MA Y J,ZHANG X Y,et al.Effects of rotation on osteonectin and osteopontin mRNA level of cultured osteoblasts in rats[J]. Acta Physiological Sinica,2005,57(3):384-388.
[24]	THURNER P J,CHEN C G,IONOVA-MARTAIN S,et al.Osteopontin deficiency increases bone fragility but preserves bone mass[J]. Bone,2010,46(6):1564-1573.
[25]	REZAL S,SHAUKAT A,MAHMUD M,et al.Evaluation of osteopontin in combination with bone turnover markers for the assessment of osteoporosis in postmenopausal women[J]. British Journal of Medicine and Medical Research,2016,14(6):1-11.
[26]	GERICKE A,QIN C,SPEVAK L,et al.Importance of phosphorylation for osteopontin regulation of biomineralization[J]. Calcified Tissue International,2005,77(1):45-54.
[27]	HUNTER G K,HAUSCHKA P V,POOLE R A,et al.Nucleation and inhibition of hydroxyapatite formation by mineralized tissue proteins[J]. Biochemical Journal,1996,317(1):59-64.
[28]	YOKOSAKI Y,HIGASHIKAWA F.Osteopontin receptors and signal transduction[J]. Nihon Rinsho,2005,63(10):613-617.

Sequence and Expression Analysis of OPN Gene in Tibia of Layer During Late Period of Laying

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