Generating antibodies to the gene 3 proteins of infectious bronchitis virus

Amanda R. Pendleton, Carolyn E Machamer

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Infectious bronchitis virus (IBV), a group 3 coronavirus, produces three proteins (IBV E, IBV 3a, and IBV 3b) from subgenomic mRNA 3 during infection. IBV E, a viral envelope protein, plays a role in virus budding, possibly by altering membrane morphology at the virus assembly site. In addition to this role, IBV E may also function as a viroporin, although no data from infected cells have confirmed this possibility definitively. Conversely, the IBV 3a and IBV 3b proteins are nonstructural proteins. These proteins are dispensable for replication in cell culture, but are thought to be important for infection of the natural host. This chapter details methods for generating and screening antibodies to these gene 3 proteins. Antibodies were raised in rabbits following inoculation with IBV-specific peptides and GST fusion proteins, and were screened by immunofluorescence, radioimmunoprecipitation, and immunoblotting.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages163-189
Number of pages27
Volume454
ISBN (Print)9781588298676
DOIs
StatePublished - 2008

Publication series

NameMethods in Molecular Biology
Volume454
ISSN (Print)10643745

Fingerprint

Infectious bronchitis virus
Antibodies
Proteins
Viral Envelope Proteins
Virus Release
Virus Assembly
Coronavirus
Infection
Immunoblotting
Fluorescent Antibody Technique
Cell Culture Techniques
Rabbits

Keywords

  • Accessory proteins
  • Antibodies
  • Coronavirus
  • Infectious bronchitis virus, 3a, 3b, E

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Pendleton, A. R., & Machamer, C. E. (2008). Generating antibodies to the gene 3 proteins of infectious bronchitis virus. In Methods in Molecular Biology (Vol. 454, pp. 163-189). (Methods in Molecular Biology; Vol. 454). Humana Press. https://doi.org/10.1007/978-1-59745-181-9_14

Generating antibodies to the gene 3 proteins of infectious bronchitis virus. / Pendleton, Amanda R.; Machamer, Carolyn E.

Methods in Molecular Biology. Vol. 454 Humana Press, 2008. p. 163-189 (Methods in Molecular Biology; Vol. 454).

Research output: Chapter in Book/Report/Conference proceedingChapter

Pendleton, AR & Machamer, CE 2008, Generating antibodies to the gene 3 proteins of infectious bronchitis virus. in Methods in Molecular Biology. vol. 454, Methods in Molecular Biology, vol. 454, Humana Press, pp. 163-189. https://doi.org/10.1007/978-1-59745-181-9_14
Pendleton AR, Machamer CE. Generating antibodies to the gene 3 proteins of infectious bronchitis virus. In Methods in Molecular Biology. Vol. 454. Humana Press. 2008. p. 163-189. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-59745-181-9_14
Pendleton, Amanda R. ; Machamer, Carolyn E. / Generating antibodies to the gene 3 proteins of infectious bronchitis virus. Methods in Molecular Biology. Vol. 454 Humana Press, 2008. pp. 163-189 (Methods in Molecular Biology).
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