EXPRESSION OF RECOMBINANT US9 PROTEIN OF BOVINE HERPESVIRUS TYPE1.1 (BOHV-1.1) IN INSECT CELL LINE AND EXAMINE ITS APPLICATION IN AN IMMUNO ASSAY FOR THE DIAGNOSIS OF BOHV-1.1 INFECTED CATTLE

Authors

  • ENGY A. OSMAN Department of Genetics, Faculty of Agriculture, Cairo University, Giza
  • W. H. ELMENOFY Agriculture Genetic Engineering Research Institute (AGERI), ARC, Giza
  • A. A. EL-KHOLY Veterinary Vaccine and Serum Research Institute (VSRI), Abbasia
  • M. H. SOLIMAN Department of Genetics, Faculty of Agriculture, Cairo University, Giza
  • LAMIAA EL-GAIED Agriculture Genetic Engineering Research Institute (AGERI), ARC, Giza
  • NAGLAA A. ABDALLAH Department of Genetics, Faculty of Agriculture, Cairo University, Giza

Abstract

In this study, a recombinant baculovirus expressing the US9 tegument protein of the Egyptian BoHV-1.1 Abu-Hammad strain (vAc-Us9) was constructed using Bac-to-Bac expression system. A coding fragment of 435bp of Us9 gene was cloned into pFastBac-1 expression vector then transferred into the baculovirus expression vector system (BEVs) for expression f BoHV-1 US9 tegument protein. The PCR analysis of the recombinant virus confirmed the successful transposition of the expression cassette into the recombinant bacmid. The expressed US9 protein in infected cell culture was assayed using Indirect Immunofluorescence Assay (IFA) in addition to SDS-PAGE and western blot using crude cell lysate. The US9 protein expressed in infected cell culture supernatant as well as cell lysate were used as a coating antigen in an indirect enzyme linked immunosorbent assay (ELISA) to examine its sensitivity for the detection of the specific antibodies that generated against BoHV-1. The effectiveness of using of the expressed US9 protein as antigen for anti-US9 detection in serum samples infected with BoHV-1 was discussed in details.

References

Abdelmagid O., Mansour M. M., Okwumabua O. and van DrunenLittle-van den Hurk S. (1998). Expression and cellular distribution of baculovirus-expressed bovine herpesvirus 1 (BHV-1) glycoprotein D (g D) sequences. Arch., 143:2173-2187.

Bertolotti C., Griffiths S. M., Truelove N. K., Box S. J., Preziosi R. F. and de Leon P. S. (2015). Isolation and characterization of 10 polymorphic microsatellite loci for the endangered Galapagos-endemic whitespotted sandbass (Paralabraxalbomaculatus). PeerJ 3:e1253; DOI 10.7717/peerj.1253.

Chandranaik B. M., Rathnamma D., Patil S. S., Ranganatha S., Ramesh C. K., Akhila D. S., Isloor S., Renukaprasad C. and Prabhudas K. (2014). Epidemiology of bovine herpes virus-1 under different housing practices in cattle breeding stations. Indian J. Anim. Sci., 84: 103-107.

Chowdhury S. I., Mahmood S., Simon J., Al-Mubarak A. and Zhou Y. (2006). The Us9 gene for bovine herpesvirus 1 (BHV-1) effectively complements a Us9-null strain of BHV-5 for anterograde transport, neurovirulence, and neurovasiveness in a rabbit model. J. Virol., 80:4396-4405.

Chowdhury S. I., Brum M.C. S., Coats C., Doster A., Wei H. and Jones C. 2011. The bovine herpesvirus type 1 envelope protein US9 acidic domain is crucial for anterograde axonal transport. Vet. Microbiol.,doi: 101016/J.Vetmic.2011.05.012.

El-Kholy A. A. and Abdelrahman K. A. 92006). Genetic characterization of the Egyptian vaccinal strain Abu-Hammad of bovine herpesvirus-1. Rev. Sci. Tech. Off. Int. Epiz., 25:1081-1095.

El-kholy A. A., Abdou E. R., Rady D. I. and Elseafy M. M. (2013). Baculovirus expression and diagnostic utility of the glycoprotein E of bovine herpesvirus-1.1 Egyptian strain ‘’ Abu-Hammad’’. J. Virol. Meth., 191:33-40.

Enquist, L. W. 2002. Exploiting circuit-specific spread of pseudorabies virus in the central nervous system: insights to pathogenesis and circuit tracers. J. Infect. Dis. 186:209-214.

Hafez, S.M., Bazz T. I., Mohsen A. Y. and Monira H. (1976). Infectious bovine rhinotracitis in Egypt: Isolation and serological identification of the virus. J. Egypt. Vet. Med. Assoc., 36:129-139.

Jaysukh K., Samiullakhan S., Basavaraj M. and Kirit B. (2018). Seroprevalence of Infectious Bovine Rhinotracheitis (BHV-1) in dairy animals with reproductive disorder Saurashtra of Gujarat, India. Int. J. Curr. Microbiol. App. Sci., 7: 1371-1376.

Kost T. A., Condreay J. P. and Jarvis D. L. (2005). Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat. Biotechnol., 23:567-575.

Laemmli U. K. (1970). Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227:680-685.

Luckow V. A. and Summers M. D. (1988). Signals important for high-level expression of foreign genes in Autographacalifornica Nuclear Polyhedrosis Virus Expression Vector. Virology, 167:56-71.

Lyman M.G., Kemp C. D., Taylor M. P. and Enquist L. W. (2009). Comparison of the pseudorabies virus US9 protein with homologs from other veterinary and human alpha herpesviruses. J. Virol., 83:6978-6986.

Massotte, D. (2003). G protein-coupled receptor overexpression with the baculovirus–insect cell system: a tool for structural and functional studies. Biochimicaet Biophysica Acta, 1610: 77-89.

Matsuura Y., Possee R. D., Overton H. A. and Bishop D. L. (1987). Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. J. Gen. Virol., 68:1233-1250.

Meurens F., Schynts F., Keil G. M., Muylkens B., Vanderplasschen A., Gallego P. and Thiry E. (2004). Superinfection prevents recombination of the alpha herpesviruses bovine herpesvirus 1. J. Virol., 78:3872-3879.

Momtaz H.and Abbasian B. (2009). Expression of infectious bovine rhinotracitis virus glycoprotein B in bacterial cell. J. of Animal and Vet. Advances, 8:1735-1739.

Murphy, C. I., Piwnica-Worms, H., Grunwald S., Romanow W. G., Francis, N. and Fan H. Y. (2004). Overview of the baculovirus expression system. In: F. M. Ausubel et al. (EDs.), Current protocols in Molecular Biology. Wiley, New York, p.16.9-16.11.

Muylkens B., Thiry J., Kirten P., Schynts F. and Thiry E. (2007). Bovine herpesvirus 1 infection and infectious bovine rhinotrachitis. Vet. Res., 38:181-209.

Robinson K.E., Meers J., Gravel J. L., McCarthy F. M. and Mahony T. J. (2008). The essential and nonessential genes of bovine herpesvirus 1. J. General Virol., 89:2851-2863.

Sambrook J. and Russell D. W. (2000). Molecular cloning: A laboratory manual. Cold Spring Harbour Laboratory, NY, USA.

Schwyzer M. and Ackermann M. (1996). Molecular virology of ruminant herpesviruses. Vet. Microbio., 53:17-29.

Tomishima M. J. and Enquist L. W., (2001). A conserved –herpesvirus protein necessary for axonal localization of viral membrane proteins. J. CellBiol., 154:741-752.

Tomishima M. J., G. A. Smith and L. W. Enquist. (2001). Sorting and transport of alpha herpesviruses in axons. Traffic, 2:429-436.

Trowitzsch S., Bieniossek C., Nie Y., Garzoni F. and Berger I. (2010). New baculovirus expression tools for recombinant protein complex production. J. Struct. Biol., 172:45-54.

Van Drunen Little-van den Hurk S., Parker M. D., Massie B., Van den Hurk J. V., Harland R., Babiuk L. A. and Zamb T.J. (1993). Protection of cattle from BHV-1 infection by immunization with recombinant glycoprotein gIV. Vaccine, 11:25-35.

Voller A., Bridwell, D. E., Bartlett A. (1976). Enzyme immunoassay in diagnostic medicine: theory and practice. Bull. World Health Organ, 53: 55-65.

Yoshitake N., Xuan, X. and Otsuka H. (1997). Identification and characterization of bovine herpesvirus-1 glycoproteins E and I. J. of General Virol., 78: 1399-1403.

Yu-chen HU. (2005). Baculovirus as a highly efficient expression vector in insect and mammalian cells. ActaPhamaco. Sinica, 26: 405-416.

Downloads

Published

2020-08-30

Issue

Vol. 49 No. 1 (2020)

Section

Articles