Related Experiment Video
Updated: Jul 15, 2026
Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
[Immunogenicity of co-expressed p30-gp90 against reticuloendotheliosis virus]
Objective:
To study the immunogenicity of co-expression of p30, one of a group specific antigens, and glycoprotein gp90 genes of Reticuloendotheliosis virus (REV).
Methods:
p30 and gp90 genes were amplified with the template of plasmid pPB101 containing the whole sequence of spleen necrosis virus (SNV), and cloned into pET-28a(+) vector. The positive clone pET-p30-gp90 was identified by enzyme analysis and sequencing. The co-expressed protein p30-gp90 was confirmed by SDS-PAGE and Western blot analysis. After quantitation, the purified protein p30-gp90 was immunized to Balb/c mice three times to prepare the p30-gp90 specific anti-serum. Afterwards, we detected the REV infected Chick Embryo Fibroblasts (CEF) by Immunofluorescence assay (IFA) with the prepared anti-serum.
Results:
The p30-gp90 was co-expressed efficiently by SDS-PAGE and Western blot analysis. The anti-serum of p30-gp90 reacts with REV infected CEF in IFA.
Conclusion:
The expressed protein of p30-gp90 keeps good immunogenicity.
Insights
The co-expression of Reticuloendotheliosis virus (REV) p30 and glycoprotein gp90 genes resulted in a protein with good immunogenicity. This protein can be used to detect REV-infected cells, aiding in disease diagnosis.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Context:
- Reticuloendotheliosis virus (REV) poses a significant threat to poultry health.
- Understanding REV gene function and developing diagnostic tools are crucial for disease control.
Purpose:
- To investigate the immunogenicity of co-expressed p30 and glycoprotein gp90 genes from REV.
- To develop a potential diagnostic marker for REV infection.
Summary:
- The p30 and gp90 genes of REV were successfully amplified, cloned, and co-expressed as a single protein (p30-gp90).
- The purified p30-gp90 protein demonstrated efficient co-expression and induced a specific immune response in mice, generating an anti-serum.
- The generated anti-serum effectively detected REV-infected Chick Embryo Fibroblasts (CEF) via Immunofluorescence Assay (IFA).
Impact:
- The study confirms the immunogenic potential of the co-expressed p30-gp90 protein.
- This finding supports the development of novel diagnostic strategies for Reticuloendotheliosis.
More Related Videos
07:10Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay
Published on: September 14, 2014
05:15Detection of Neutralization-sensitive Epitopes in Antigens Displayed on Virus-Like Particle VLP-Based Vaccines Using a Capture Assay
Published on: February 10, 2022
Related Concept Videos
01:29Immune Response Against Viral Pathogens
NK Cells
NK cells are a crucial part of our innate immune system, acting as the first line of defense against viral infections. These cells can recognize and kill infected cells without prior exposure to the virus, effectively slowing down the spread of infection. Additionally, NK cells produce proinflammatory...
01:25Immunodeficiency Diseases
There are three main causes of immunodeficiency disorders...