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Expression and purification of recombinant Marek's disease virus serotype 1 specific phosphorylated protein pp38 in
1Department of Radiation Biology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Abstract:
Phosphorylated protein pp38 is the only protein that is detected in the Marek's disease (MD) lymphoma caused by MD virus serotype 1 (MDV-1) and lymphoblastoid cell lines. In this study, a recombinant protein coded for by the almost entire open reading frame of the MDV-1 pp38 cDNA was produced in E. coli and purified by affinity chromatography. Prior to the expression and purification of the protein, cDNA containing the entire coding region for pp38 was cloned and its nucleotide sequence was determined. Immunoblot analysis indicated that the expressed recombinant protein electrophoresed close to that of the pp38 in infected cells. The difference in mobility of the purified recombinant and the pp38 in infected cells corresponded to a fusion peptide. The recombinant pp38 may be of interest for function analyses and the diagnotic use of pp38.
Insights
Researchers produced a recombinant Marek's disease virus pp38 protein for diagnostic use. This purified protein aids in understanding Marek's disease lymphoma and offers potential diagnostic applications.
Area of Science:
- Veterinary Virology
- Molecular Biology
- Immunology
Background:
- Marek's disease (MD) lymphoma is characterized by the presence of phosphorylated protein pp38, exclusively associated with MD virus serotype 1 (MDV-1).
- This protein is also detected in lymphoblastoid cell lines, indicating its significance in MDV-1 pathogenesis.
Purpose of the Study:
- To produce and purify a recombinant pp38 protein from MDV-1.
- To evaluate the characteristics of the recombinant pp38 protein for potential diagnostic and functional analyses.
Main Methods:
- Cloning of the MDV-1 pp38 cDNA and determination of its nucleotide sequence.
- Expression of the recombinant pp38 protein in E. coli.
- Purification of the recombinant protein using affinity chromatography.
- Immunoblot analysis to compare the expressed protein with pp38 from infected cells.
Main Results:
- A recombinant protein, nearly identical to the MDV-1 pp38, was successfully expressed and purified.
- Immunoblot analysis confirmed the expressed protein's similarity to pp38 in infected cells, with a slight mobility difference attributed to a fusion peptide.
- The nucleotide sequence of the pp38 cDNA was determined prior to expression.
Conclusions:
- The successful production of recombinant pp38 protein provides a valuable tool for further research.
- The recombinant pp38 protein shows potential for use in functional studies and diagnostic applications for Marek's disease.