Comparison of three methods in construction fusion gene of influenza A virus Nucleoprotein / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 70-74, 2012.
Artigo
em Chinês
| WPRIM
| ID: wpr-246177
ABSTRACT
<p><b>OBJECTIVE</b>To develop a best method of constructing influenza NP fusion gene containing enhanced green fluorescent protein (EGFP).</p><p><b>METHODS</b>The full-length NP gene of influenza A was amplified by RT-PCR and was inserted into an eukaryotic expression vector pEGFP-N1 in order to construct a fusion gene of pEGFP-N1-NP using three different methods. Method one, NP gene containing restriction endonucleases and pEGFP-N1 were both digested using the same restrict enzymes and ligated, yielding the fusion gene of pEGFP-N1-NP. Method two, NP gene was cloned into pMD19-T Vector to construct a plasmid of pMD19-T-NP. The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP; Method three, NP gene containing restriction endonucleases was cloned into pMD19-T Simple Vector to construct a plasmid of pMD19-T-NP. The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP.</p><p><b>RESULTS</b>The fusion gene of recombinant eukaryotic expression vector pEGFP-N1-NP was successfully constructed by using method three.</p><p><b>CONCLUSIONS</b>The full-length NP gene is obtained and its fusion gene of recombinant eukaryotic expression plasmid is successfully constructed. This study provides foundation for further understanding the biological function of NP protein and the mechanism of diseases induced by influenza A virus.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas do Core Viral
/
Reação em Cadeia da Polimerase
/
Proteínas de Ligação a RNA
/
Clonagem Molecular
/
Fusão Gênica Artificial
/
Proteínas de Fluorescência Verde
/
Vetores Genéticos
/
Genética
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2012
Tipo de documento:
Artigo
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