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Data/hora: 09/06/2017 09:47:16
Provedor de dados: 101
País: Cuba
Título: MOLECULAR CLONING AND EXPRESSION OF A FRAGMENT OF THE GENE
CODIFYING FOR THE PROTEIN ERNS OF CLASSICAL SWINE FEVER VIRUS
Autores: Agüero,J.A; Sánchez,O; Barrera,Maritza; Toledo,J.R.
Data: 2008-08-01
Ano: 2008
Palavras-chave: Erns; CSFV; Recombinant protein; IMAC.
Resumo: Classical swine fever virus (CSFV), belonging to the genus Pestivirus of the Flaviviridae
family, is an enveloped positive stranded RNA virus highly contagious that can cause a
fatal disease, characterized by fever, leukopenia and hemorrhage, with substantial
economic losses. There is a great demand for a marker vaccine against CSFV. C, Erns,
E1, and E2 are the structural proteins of the virus. E2 is the best candidate to be
incorporated in vaccine, while Erns becomes an ideal candidate as an antigen in a
differential diagnostic test. A synthetic fragment of the Erns gene (codifying for aa
109-160) was subcloned into pET28a vector. The cloning was screened by restriction
analysis. The gene was expressed as a his-tag fusion protein in BL21 (DE3) E. coli strain.
The recombinant polypeptide formed aggregates of about 7.9; 15.8; 23.7; 31.6; 39.5; 47.5
and 55.3kDa. The protein was not recognized by Western blot using an antibody against
the virus. Using an ion metal affinity chromatography procedure, a 90% pure recombinant
product was obtained. The potential use of this antigen for detection of CSFV antibodies
should be further evaluated.
Tipo: Journal article
Idioma: Inglês
Identificador: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S0253-570X2008000200003
Editor: Centro Nacional de Sanidad Agropecuaria
Formato: text/html
Fonte: Revista de Salud Animal v.30 n.2 2008