羊口疮病毒ORF125基因编码蛋白的结构预测及其在真核表达质粒转染细胞中的定位

doi:10.11843/j.issn.0366-6964.2014.08.025

Abstract

Abstract:

The current study was undertaken to forecast the secondary structure and find out the localization of transfected cells of ORF125 from orf virus (ORFV).ORF125 gene was acquired by PCR and cloned into pEGFP-N1 eukaryotic expression vector directionally.The recombinant expression plasmid was identified by restriction enzyme digestion analysis，sequencing and named pEGFP-ORF125.Then the phylogenetic analysis of the gene was made by DNAStar and MEGA4.0.Also the secondary structure of ORF125 was predicted and compared with cellular Bcl-2 proteins by Expert Protein Analysis System (EXPASY).The recombinant plasmid was transfect into goat skin fibroblasts (GSFs)，subcellular localization of ORF125 was examined with laser con-focal microscopy after DAPI staining.The results of sequencing alignment showed that the homology of China Luoyang 2013 shared 97.1%-98.1% with other ORFV isolates and 84.1%（GQ329669）and 85.6%（GQ329670）with another two Pseudocowpox isolates from different regions at nucleotides level respectively.The sequence identity of ORF125 and cellular Bcl-2 proteins is low，but the secondary and tertiary structures are conserved.Laser con-focal microscopy results showed that ORF125 distributed dispersively in the cytoplasm of GSFs.This study helped to further understanding the function of ORF125.Results verified the ORFV ORF125 gene is conserved and have similar function with Bcl-2 protein，in addition，the ORF125 gene could be expressed successfully in GSFs，and the ORF125 protein localizes in the cytoplasm.

CLC Number:

S852.659.1

KONG Han-jin，ZHANG Ke-shan，LIU Yong-jie，SHANG You-jun，LIU Xiang-tao，WU Bin. Structure Prediction and Localization of ORF125 from Orf Virus on Transfected Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(8): 1375-1380.

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References

［1］BÜTTNER M，RZIHA H J.Parapoxviruses：from the lesion to the viral genome［J］.J Vet Med B Infect Dis Vet Public Health，2002，49(1):7-16.
［2］MAZUR C，FERREIRA I I，RANGEL FILHO F B，et al.Molecular characterization of Brazilian isolates of orf virus［J］. Vet Microbiol，2000，73(4):253-259.
［3］COTTONE R，BÜTTNER M，BAUER B，et al.Analysis of genomic rearrangement and subsequent gene deletion of the attenuated Orf virus strain D1701［J］.Virus Res，1998，56(1):53-67.
［4］WESTPHAL D，LEDGERWOOD E C，HIBMA M H，et al.A novel Bcl-2-like inhibitor of apoptosis is encoded by the parapoxvirus ORF virus［J］.J Virol，2007，81(13):7178-7188.
［5］WESTPHAL D，LEDGERWOOD E C，TYNDALL J D，et al.The orf virus inhibitor of apoptosis functions in a Bcl-2-like manner，binding and neutralizing a set of BH3-only proteins and active Bax［J］.Apoptosis，2009，14(11):1317-1330.
［6］GAO Y，SHAO S，XIAO X，et al.Using pseudo amino acid composition to predict protein subcellular location：approached with Lyapunov index，Bessel function，and Chebyshev filter［J］.Amino Acids，2005，28(4):373-376.
［7］YOULE R J，STRASSER A.The BCL-2 protein family：opposing activities that mediate cell death［J］.Nat Rev Mol Cell Biol，2008，9(1):47-59.
［8］EVERETT H，MCFADDEN G.Apoptosis：an innate immune response to virus infection［J］.Trends Microbiol，1999，7(4):160-165.
［9］CUCONATI A，WHITE E.Viral homologs of BCL-2：role of apoptosis in the regulation of virus infection［J］.Genes Dev，2002，16(19):2465-2478.
［10］HARDWICK J M，BELLOWS D S.Viral versus cellular BCL-2 proteins［J］.Cell Death Differ，2003，10(Suppl 1):S68-S76.
［11］TULMAN E R，AFONSO C L，LU Z，et al.The genome of canarypox virus［J］.J Virol，2004，78(1):353-366.
［12］EVERETT H，BARRY M，LEE S F，et al.M11L：a novel mitochondria-localized protein of myxoma virus that blocks apoptosis of infected leukocytes ［J］.J Exp Med，2000，191(9):1487-1498.
［13］WASILENKO S T，STEWART T L，MEYERS A F，et al.Vaccinia virus encodes a previously uncharacterized mitochondrial-associated inhibitor of apoptosis［J］.Proc Natl Acad Sci U S A，2003，100(24):14345-14350.
［14］ZHANG K，SHANG Y，JIN Y，et al.Diagnosis and phylogenetic analysis of Orf virus from goats in China：a case report［J/OL］.Virol J，2010，7：78.［2014-5-27］.http://www.virologyj.com/content/7/1/78.

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Structure Prediction and Localization of ORF125 from Orf Virus on Transfected Cells

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