绵羊GPx5基因克隆、生物信息学分析及表达特性的研究

doi:10.11843/j.issn.0366-6964.2014.07.007

Abstract

Abstract:

The aim of this study was to clone ovine GPx5 (Selenium-independent glutathione peroxidase 5)，and predict the structure and function of GPx5 protein，and investigate its expression and location characteristics in ovine testis and epididymis caput.The ovine GPx5 was cloned by 5′RACE and RT-PCR.The structure，characteristics and function of ovine GPx5 protein were analyzed by bioinformatics methods.The expression characteristics were detected by immunohistochemistry assays in ovine testis and epididymis caput，as well as by Western blotting assays in seminal plasma.The results showed GPx5 gene (GenBank accession No.：JQ 320282) contained an 660 bp open reading frame，which coded 219aa with molecular weight of 25 ku.Its potential signal peptide was 1-21aa.The domain structure of tertiary structure was alpha/beta structure.There were 11 potentail phosphorylation sites predicted and 7 conserved and specific phosphokinase binding sites.The prediction results of InterProScan showed 38-151aa were the conserved protein domain of GSHPx family and 36-121aa was the conserved protein domain of thioredoxin-like superfamily.GPx5 protein was essentially presented in pseudostratified columnar epithelium，and poorly presented in epithelium of the ovine epididymis and Leydig cell of ovine testis，and also presented in fluid of epididymis caput and seminal plasma in rams.Ovine GPx5 protein was secreted，which had a GPx activity and function of thioredoxin-like.GPx5 could be as a mediator of signaling pathway and need be tested by further research.

CLC Number:

S826
S813.3

ZHANG Chun-xiang，GUO Yun-yan，BAI Yuan-sheng,YANG Zi-sen，ZHANG Guo-lin，JIAO Guang-yue，REN You-she. Clone，Bioinformatics Analysis and Expression Characteristics of Ovine GPx5[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(7): 1075-1083.

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References

［1］PERRY A C，JONES R，NIANG L S，et al.Genetic evidence for an androgen-regulated epdiidymal secretory glutathione peroxidase whose transcript does not contain a selenocysteine codon［J］.Biochem J，1992，285：863-870.
［2］VERNET P，RIGAUDIERE N，GHYSELINCK N，et al.In vitro expression of a mouse tissue specific glutathione-peroxidase like protein lacking the selenocysteine can protect stably transfected mammalian cells against oxidative damage［J］.Biochem Cell Biol，1996，74：125-131.
［3］HERBETTE S，ROECKEL P，DREVET J R，et al.Seleno-independent glutathione peroxidases.More than simple antioxidant scavengers［J］.FEBS J，2007，274(9):2163-2180.
［4］SELLES B，HUGO M，TRUJILLO M，et al.Hydroperoxide and peroxynitrite reductase activity of poplar thioredoxin dependent glutathione peroxidase 5：kinetics，catalytic mechanism and oxidative inactivation［J］.Biochem J，2012，442(2):369-380.
［5］REJRAJI H，VERNET P，DREVET J R，et al.GPX5 is present in the mouse caput and cauda epididymidis lumen at three different locations［J］.Mol Reprod Dev，2002，63(1):96-103.
［6］TAYLOR A，ROBSON A，HOUGHTON B C，et al.Epididymal specific，selenium-independent GPX5 protects cells from oxidative stress-induced lipid peroxidation and DNA mutation［J］.Hum Reprod，2013，28(9):2332-2342.
［7］CHABORY E，DAMON C，LENOIR A，et al.Epididymis seleno-independent glutathione peroxidase 5 maintains sperm DNA integrity in mice［J］.J Clin Invest，2009，119(7):2074-2085.
［8］CHABORY E，DAMON C，LENOIR A，et al.Mammalian glutathione peroxidases control acquisition and maintenance of spermatozoa integrity［J］.J Anim Sci，2010，88(4):1321-1331.
［9］AITKEN R J.Gpx5 protects the family jewels［J］.J Clin Invest，2009，119(7):1849-1851.
［10］GHYSELINK N B，DUFAURE J P.A mouse cDNA sequence for epididymal androgen-regulated proteins related to glutathione peroxidase［J］.Nucleic Acids Res，1990，18(23):7144.
［11］WILLIAMS K，FRAYNE J，HALL L.Expression of extracellular glutathione peroxidase type 5 (GPX-5) in the rat male reproductive tract［J］.Mol Hum Reprod，1998，4：841-848.
［12］SCHWAAB V，LAREYRE J J，VERNET P，et al.Characterization，regulation of the expression and putative roles of two glutathione peroxidase proteins found in the mouse epididymis［J］.J Reprod Fertil，1998,53S:157-162.
［13］ZHANG T，CHABORY E，BRITAN A，et al.GPX5，the selenium-independent glutathione peroxidase-encoding single copy gene is differentially expressed in mouse epididymis［J］.Reprod Fertil Dev，2008，20(5):615-625.
［14］GUYONNET B，MAROT G，DACHEUX J L，et al.The adult boar testicular and epididymal transcriptomes［J］.BMC Genomics，2009，10：369-398.
［15］MANASKOVA P，DAVIDOVA N，JONAKOVA V.Biochemical and bingding characteristics of boar epididymal fluid proteins［J］. J Chromato，2011，879(1):100-106.
［16］NOVAK S，RUIZ A，DIXON W T，et al.Seminal plasma proteins as potential markers of relative fertility in boars［J］.J Androl，2010，31(2):188-200.
［17］张建新，王茜，荀文娟，等.GPx家族基因在山羊不同组织和睾丸不同发育时期的表达特性研究［J］.畜牧兽医学报，2011，42(5)：650-657.
［18］GRIGNARD E，MORIN J，VERNET P，et al.GPX5 orthologs of the mouse epididymis-restricted and sperm-bound selenium-independent glutathione peroxidase are not expressed with the same quantitative and spatial characteristics in large domestic animals［J］.Theriogenology，2005，64(4):1016-1033.
［19］ARNOLD K，BORDOLI L，KOPP J，et al.The SWISS-MODEL Workspace：A web-based environment for protein structure homology modeling［J］.Bioinformatics，2006，22：195-201.
［20］ZDOBNOV E M，APWEILER R.InterProScan-an integration platform for the signature-recognition methods in InterPro［J］.Bioinformatics，2001，17(9):847-848.
［21］MARCHLER B A，LU S，ANDERSON J B，et al.CDD：a Conserved Domain Database for the functional annotation of proteins［J］.Nucleic Acids Res，2011，39(Database issue):D225-229.
［22］COOPER T G，YEUNG C H，WAGENFELD A，et al.Mouse models of infertility due to swollen spermatozoa［J］.Mol Cell Endocrinol，2004，216(1-2):55-63.
［23］http://www.uniport.org/uniport/O75715.
［24］HALL L，WILLIAMS K，PERRY A C，et al.The majority of human glutathione peroxidase type 5 (GPX-5) transcripts are incorrectly spliced：implications for the role of GPX-5 in the male reproductive tract ［J］.J Biochem，1998，333：5-9.

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Clone，Bioinformatics Analysis and Expression Characteristics of Ovine GPx5

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