Purification of the Sendai virus nonstructural C protein expressed in E. coli, and preparation of antiserum against C protein.
An expression plasmid, ptac-C, was constructed by inserting the cDNA of the coding region of the Sendai virus nonstructural C protein downstream of the tac promoter of E. coli expression plasmid ptac12-Bam. A new protein produced in E. coli after induction was purified to near homogeneity. The purified protein was found to be identical with the C protein predicted from the C gene cDNA in molecular weight, isoelectric point, amino acid composition, and the amino acid sequence at the N-terminal of the protein as well as those of several fragments obtained on V8 protease digestion. Antiserum raised against the purified protein specifically reacted with the C protein in infected cells. Using this antiserum, the localization of the C protein in infected cells was examined by immunofluorescence, which revealed that it appeared in the cytoplasm but not in nuclei.
Omata-Yamada
T
T
Department of Physiological Chemistry, Tokyo Metropolitan Institute of Medical Science, Japan.
Hagiwara
K
K
Katoh
K
K
Yamada
H
H
Iwasaki
K
K
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D051379:Mice; D010222:Parainfluenza Virus 1, Human / Q000235:genetics / Q000302:isolation & purification; D010957:Plasmids; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C051221:nonstructural C protein, Sendai virus