Expression of infectious laryngotracheitis virus glycoproteins in Escherichia coli and their application in enzyme-linked immunosorbent assay.
Three glycoproteins of infectious laryngotracheitis virus (ILTV), gC, gE, and gp60, were expressed in Escherichia coli as fusion proteins with a 6-histidine tag at their amino termini. The proteins expressed, designated as r-gC, r-gp60, and r-gE, all retain their antigenicity, as revealed by Western blot with chicken antiserum against ILTV. However, only r-gp60 and r-gE, but not r-gC, were found to be soluble. The soluble r-gp60 and r-gE were purified by a nickel column and then used as the enzyme-linked immunosorbent assay (ELISA) antigen for detecting ILTV-specific antibodies. The diagnostic potential of r-gE and r-gp60 ELISA was assessed with the use of sera prepared from vaccinated or unvaccinated chickens of either specific-pathogen-free (SPF) or field origins. The result shows that r-gp60 and r-gE ELISA could discriminate vaccinated SPF chickens from unvaccinated ones 2 wk postvaccination. Moreover, r-gp60 and r-gE ELISA could also discriminate vaccinated field flocks from unvaccinated ones. This result indicates that r-gp60 and r-gE might serve as an alternative ELISA antigen for detecting ILTV-specific antibodies. Moreover, r-gp60 or r-gE ELISA might play an important role in the eradication of infectious laryngotracheitis (ILT) in the future when the gp60- or gE-deleted marker vaccine of ILT is available.
Chang
Poa-Chun
PC
Institute of Veterinary Microbiology, National Chung Hsing University, Taichung, Taiwan.
Chen
Kwan-Ting
KT
Shien
Jui-Hung
JH
Shieh
Happy K
HK
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western / Q000662:veterinary; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D004926:Escherichia coli; D015967:Gene Expression Regulation, Viral; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D001352:Herpesvirus 1, Gallid / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D012995:Solubility; D013047:Specific Pathogen-Free Organisms; D014611:Vaccination / Q000662:veterinary
D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D011993:Recombinant Fusion Proteins