Biotechnology Letters

, Volume 29, Issue 6, pp 871–875 | Cite as

Expression of truncated phosphoproteins of Nipah virus and Hendra virus in Escherichia coli for the differentiation of henipavirus infections

  • Ji-Ming ChenEmail author
  • Koon Chu Yaiw
  • Meng Yu
  • Lin-Fa Wang
  • Qing-Hua Wang
  • Gary Crameri
  • Zhi-Liang Wang
Original Research Paper


The genus Henipavirus in the family Paramyxoviridae compromises two newly identified dangerous pathogens, Nipah virus and Hendra virus. Phosphoprotein of the two viruses is one of the major immunodominant antigens and the most divergent protein in the viral genomes. We have now expressed two pairs of truncated phosphoproteins of the two viruses in Escherichia coli in a soluble form using a vector tailored from pET32a. The truncated recombinant phosphoproteins were purified with His-Tag affinity chromatography and their antigenicity was determined by western blotting and ELISA. The longer pair of truncated recombinant phosphoproteins, covering amino acid residues 4–550, was more antigenic than the shorter one and of potential utility in the serological differentiation of henipavirus infections.


Escherichia coli Hendra virus Nipah virus Phosphoprotein Serological differentiation Viral infections 



This work was supported in part by grants from Chinese Ministry of Science & Technology (No. 2003BA712A04-04) and Australian Biosecurity Cooperative Research Center for Emerging Infectious Diseases (Project 1.013RE).


  1. Chadha MS, Comer JA, Lowe L, Rota PA, Rollin PE, Bellini WJ, Ksiazek TG, Mishra A (2006) Nipah virus-associated encephalitis outbreak, Siliguri, India. Emerg Infect Dis 12:235–240PubMedGoogle Scholar
  2. Chan YP, Koh CL, Lam SK, Wang LF (2004) Mapping of domains responsible for nucleocapsid protein-phosphoprotein interaction of Henipaviruses. J Gen Virol 85:1675–1684PubMedCrossRefGoogle Scholar
  3. Chen JM, Yu M, Morrissy C, Zhao YG, Meehan G, Sun YX, Wang QH, Zhang W, Wang LF, Wang ZL (2006a) A comparative indirect ELISA for the detection of henipavirus antibodies based on a recombinant nucleocapsid protein expressed in Escherichia coli. J Virol Methods 136:273–276CrossRefGoogle Scholar
  4. Chen JM, Guo LX, Sun CY, Sun YX, Chen JW, Li L, Wang ZL (2006b) A stable and differentiable RNA positive control for reverse transcription-polymerase chain reaction. Biotechnol Lett (in press)Google Scholar
  5. Choi C (2004) Nipah’s return. The lethal “flying fox” virus may spread between people. Sci Am 291:21–22CrossRefGoogle Scholar
  6. Chua KB (2003) Nipah virus outbreak in Malaysia. J Clin Virol 26:265–275PubMedCrossRefGoogle Scholar
  7. Chua KB, Bellini WJ, Rota PA, Harcourt BH, Tamin A, Lam SK, Ksiazek TG, Rollin PE, Zaki SR, Shieh W, others (2000) Nipah virus: a recently emergent deadly paramyxovirus. Science 288:1432–1435PubMedCrossRefGoogle Scholar
  8. Crameri G, Wang LF, Morrissy C, White J, Eaton BT (2002) A rapid immune plaque assay for the detection of Hendra and Nipah viruses and anti-virus antibodies. J Virol Methods 99:41–51PubMedCrossRefGoogle Scholar
  9. Daniels P, Ksiazek T, Eaton BT (2001) Laboratory diagnosis of Nipah and Hendra virus infections. Microbes Infect 3:289–295PubMedCrossRefGoogle Scholar
  10. Eaton BT, Broder CC, Wang LF (2005) Hendra and Nipah viruses: pathogenesis and therapeutics. Curr Mol Med 5:805–816PubMedCrossRefGoogle Scholar
  11. Field HE, Barratt PC, Hughes RJ, Shield J, Sullivan ND (2000) A fatal case of Hendra virus infection in a horse in north Queensland: clinical and epidemiological features. Aust Vet J 78:279–280PubMedGoogle Scholar
  12. Harcourt BH, Lowe L, Tamin A, Liu X, Bankamp B, Bowden N, Rollin PE, Comer JA, Ksiazek TG, Hossain MJ, others (2005) Genetic characterization of Nipah virus, Bangladesh, 2004. Emerg Infect Dis 11:1594–1597PubMedGoogle Scholar
  13. Hsu VP, Hossain MJ, Parashar UD, Ali MM, Ksiazek TG, Kuzmin I, Niezgoda M, Rupprecht C, Bresee J, Breiman RF (2004) Nipah virus encephalitis reemergence, Bangladesh. Emerg Infect Dis 10:2082–2087PubMedGoogle Scholar
  14. Hyatt AD, Zaki SR, Goldsmith CS, Wise TG, Hengstberger SG (2001) Ultrastructure of Hendra virus and Nipah virus within cultured cells and host animals. Microbes Infect 3:297–306PubMedCrossRefGoogle Scholar
  15. Lam SK (2003) Nipah virus––a potential agent of bioterrorism? Antiviral Res 57:113–119PubMedCrossRefGoogle Scholar
  16. Murray K, Selleck P, Hooper P, Hyatt A, Gould A, Gleeson L, Westbury H, Hiley L, Selvey L, Rodwell B, others (1995) A morbillivirus that caused fatal disease in horses and humans. Science 268:94–97PubMedCrossRefGoogle Scholar
  17. Reynes JM, Counor D, Ong S, Faure C, Seng V, Molia S, Walston J, Georges-Courbot MC, Deubel V, Sarthou JL (2005) Nipah virus in Lyle’s flying foxes, Cambodia. Emerg Infect Dis 11:1042–1047PubMedGoogle Scholar
  18. Wang LF, Harcourt BH, Yu M, Tamin A, Rota PA, Bellini WJ, Eaton BT (2001) Molecular biology of Hendra and Nipah viruses. Microbes Infect 3:279–287PubMedCrossRefGoogle Scholar
  19. Wang LF, Yu M, Hansson E, Pritchard LI, Shiell B, Michalski WP, Eaton BT (2000) The exceptionally large genome of Hendra virus: support for creation of a new genus within the family Paramyxoviridae. J Virol 74:9972–9979PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Ji-Ming Chen
    • 1
    Email author
  • Koon Chu Yaiw
    • 2
  • Meng Yu
    • 3
  • Lin-Fa Wang
    • 3
  • Qing-Hua Wang
    • 1
  • Gary Crameri
    • 3
  • Zhi-Liang Wang
    • 1
  1. 1.China Animal Health and Epidemiology CenterQingdaoChina
  2. 2.Faculty of Medicine, University of MalayaKuala LumpurMalaysia
  3. 3.Australian Animal Health LaboratoryGeelongAustralia

Personalised recommendations