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Herpes Simplex Virus Type 1 ICP27 Protein:Its Expression, Purification and Specific Antiserum Production

Herpes Simplex Virus Type 1 ICP27 Protein:Its Expression, Purification and Specific Antiserum Production
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摘要 Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid (Ni 2+ -NTA) affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection. Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid (Ni^2+-NTA) affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.
出处 《Virologica Sinica》 SCIE CAS CSCD 2010年第3期199-205,共7页 中国病毒学(英文版)
基金 The startup fund of the hundred talents program of the Chinese academy of science(20071010- 141) National natural science foundation of China (30870120) Open research fund program of the state key laboratory of virology of China (2007003, 2009007) Hubei province natural science foundation of innovation groups project (2008CDA013) Major state basic research development program (973 Program) of China (2010CB 530105)
关键词 单纯疱疹病毒 抗血清 蛋白质 纯化 生产 BLOT分析 感染细胞 P27基因 Herpes simplex virus type 1 (HSV-1) Infected-cell protein 27 (ICP27) Recombinant protein Antiserum Immunofluorescence assay.
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参考文献18

  • 1Arnau J, Lauritzen C, Petersen G E, et al. 2006. Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins. Protein Expr Purif, 48 (1): 1-13.
  • 2Brown C R, Nakamura M S, Mosca J D, et aL 1995. Herpes-Simplex Virus Transregulatory Protein Icp27 Stabilizes and Binds to 3'-Ends of Labile Messenger-Rna. J Virol, 69 (11): 7187-7195.
  • 3Fontaine-Rodriguez E C, and Knipe D M. 2008. Herpes simplex virus ICP27 increases translation of a subset of viral late mRNAs. J Virol, 82 (7): 3538-3545.
  • 4Hardy W R, and Sandri-Goldin R M. 1994. Herpes simplex virus inhibits host cell splicing, and regulatory protein ICP27 is required for this effect. J Virol, 68 (12): 7790-7799.
  • 5Hunt I. 2005. From gene to protein: a review of new and enabling technologies for multi-parallel protein expression. Protein Expr Pnrif, 40 (1): 1-22.
  • 6Johnson K E, Song B, and Knipe D M. 2008. Role for herpes simplex virus 1 ICP27 in the inhibition of type I interferon signaling. Virology, 374 (2): 487-494.
  • 7Koffa M D, Clemeats J B, Izaurralde E, et al. 2001. Herpes simplex virus ICP27 protein provides viral mRNAs with access to the cellular mRNA export pathway. EMBO J, 20 (20): 5769-5778.
  • 8Markovitz N S. 2007. The herpes simplex virus type 1 UL3 transcript starts within the UL3 open reading frame and encodes a 224-amino-acid protein. J Virol, 81 (19): 10524-10531.
  • 9Mayer M, and Buehner J. 2004. Refolding of inclusion body proteins. Methods Mol Med, 94239-94254.
  • 10McGeoch D J, Dalrymple M A, Davison A J, et al. 1988. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol, 69 ( Pt 7): 1531-1574.

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