Expression and purification of recombinant Marek's disease virus serotype 1 specific phosphorylated protein pp38 in E. coli.
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.
Endoh
D
D
Department of Radiation Biology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Niikura
M
M
Hirai
K
K
Inagaki
N
N
Hayashi
M
M
Nakajima
K
K
Sato
F
F
D000595:Amino Acid Sequence; D000956:Antigens, Viral; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D015870:Gene Expression; D015723:Gene Library; D008381:Herpesvirus 2, Gallid / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010750:Phosphoproteins / Q000096:biosynthesis*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012703:Serotyping; D016385:TATA Box; D014764:Viral Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D000956:Antigens, Viral; D017931:DNA Primers; D018076:DNA, Complementary; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C071698:phosphoprotein pp38, Marek's disease virus