Expression and purification of a neuropeptide nocistatin using two related plant viral vectors.
Gene
; 289(1-2): 69-79, 2002 May 01.
Article
em En
| MEDLINE
| ID: mdl-12036585
ABSTRACT
Both odontoglossum ringspot virus (ORSV) and tobacco mosaic virus (TMV) were investigated as expression viral vectors for the expression of a neuropeptide nocistatin. Chimeras of ORSV and TMV were constructed by fusion of 17 amino acids of mouse nocistatin (mNST) to the C-terminal of the coat protein (CP) gene via a Factor Xa cleavage linker to yield ORSV-mNST and TMV-mNST. Expression of the mNST peptide was demonstrated by immuno-transmission electron microscopy, western blot, mass spectrometry and radioimmunoassay. Serial passaging of the chimeric viruses revealed loss of mNST from TMV-mNST by the fifth passage. The mNST was maintained in ORSV-mNST throughout six passages. The mNST peptide could be effectively cleaved and purified from chimeric ORSV CP. To our knowledge, this is the first successful attempt in obtaining a complete peptide with no additional amino acid sequence after expression and purification through the use of either ORSV or TMV as vectors.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Vírus de Plantas
/
Peptídeos Opioides
/
Vetores Genéticos
Limite:
Animals
Idioma:
En
Revista:
Gene
Ano de publicação:
2002
Tipo de documento:
Article