Cloning and expression of chicken anemia virus VP3 protein / Clonagem e expressão da proteína VP3 do vírus da anemia infecciosa das galinhas (CAV)

AUTOR(ES)

Eliana Ottati Nogueira Dantas

DATA DE PUBLICAÇÃO

2007

RESUMO

Purification of chicken anemia virus (CAV) VP3 protein, expressed in a prokaryotic expression system as histidine-tagged fusion protein is demonstrated in the present study. CAV particle was obtained from infected liver of chicken and DNA was extracted. The VP3 protein gene was amplified from the extracted DNA by polymerase chain reaction (PCR) and cloned. The recombinant expression construct (pTrc-VP3) was identified by PCR and sequencing analysis. Expression of VP3 protein with a molecular mass of approximately 21 KDa was confirmed by Western blotting analysis with CAV-specific antibodies. The in vitro expressed VP3 protein was purified to near homogeneity by elution from the gel, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The purified VP3 protein was recognized by CAV antibodies in a Western blotting assay. This finding indicates that recombinant VP3 expressed in the pTrcHis2 vector system can be used as antigen to detect anti-CAV antibodies.

ASSUNTO(S)

cloning cav proteína vp3 ptrchis2 ptrchis2 cav clonagem vp3 protein

Documentos Relacionados

• Anticorpos Para o Vírus da Anemia das Galinhas (CAV) em Matrizes de Corte no Brasil
• Prevalência de anticorpos contra o vírus da anemia das galinhas (CAV) em matrizes de corte no Sul do Brasil
• Teste de reação em cadeia de polimerase dos genes VP3 e VP1 para detecção do vírus da anemia das galinhas em amostras de campo
• Detecção do vírus da anemia infecciosa das galinhas em Minas Gerais
• Vírus da anemia infecciosa das galinhas:: perfil sorológico em matrizes pesadas e presença do vírus em sua progênie.