Nucleotide sequence of the zucchini yellow mosaic virus capsid-encoding gene and its expression in Escherichia coli.
Zucchini yellow mosaic virus (ZYMV) RNA was purified and used as a template for the synthesis of cDNA. A partial restriction map covering 9.4 kb of the ZYMV genome was constructed from three clones designated ZYKS-22, ZYKS-16 and ZYKS-3. Sequencing the 3'-end region of the ZYMV genome indicates the presence of (A)48 chain. This is followed by an untranslated region of 210 nucleotides (nt) and a coding region of 837 nt corresponding to the putative virus coat protein (Cp) gene (cp). The predicted amino acid (aa) sequence of Cp derived from the cDNA showed about 50% to 62% homology with the known aa sequence for Cp of six other potyviruses. A construct of the putative cp was subcloned in frame with the lacZp gene promoter in a Bluescript plasmid and expressed in Escherichia coli cells. The fusion polypeptides (34 and 41 kDa), positively reacted in Western blots with an antiserum prepared against the native virus Cp.
Department of Virology, ARO, Volcani Center, Bet Dagan, Israel.
A
A
Gal-On
Y
Y
Antignus
A
A
Rosner
B
B
Raccah
D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D012367:RNA, Viral