Construction of a chimeric viral gene expressing plum pox virus coat protein.
Gene
; 120(2): 167-73, 1992 Oct 21.
Article
em En
| MEDLINE
| ID: mdl-1398133
ABSTRACT
The capsid-encoding gene of plum pox virus (PPV) was fused with the leader sequence of the coat protein mRNA (cp) of tobacco mosaic virus by a novel mutagenesis technique which involves reverse transcription of minus-strand RNA [synthesized by in vitro transcription of a double-stranded (ds) cDNA clone], using an ad hoc synthetic oligodeoxynucleotide as primer. The resulting cDNA was rendered ds and cloned into the plasmid, pBluescribe M13+. Transcription of this chimeric construction produced RNA molecules of 1250 nucleotides in length, which were used as messengers in the in vitro protein-synthesizing systems. The major product of this transcript consists of a 36-kDa polypeptide and was identified as the PPV coat protein (CP) by molecular weight estimation and by immunoprecipitation with a polyclonal antiserum to PPV. Transfer of this cDNA via Agrobacterium tumefaciens into plants was successfully performed. Transgenic Nicotiana plants producing the PPV CP were subsequently obtained.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Vírus de Plantas
/
Capsídeo
/
Genes Virais
Idioma:
En
Ano de publicação:
1992
Tipo de documento:
Article