Expression, structure and antigenicity analysis of N51 derived from the N-terminal heptad repeat domain in gp41 of HIV-1 CRF07_BC strain / 南方医科大学学报
Journal of Southern Medical University
; (12): 1737-1740, 2012.
Article
en Zh
| WPRIM
| ID: wpr-352344
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To express N51 derived from the N-terminal heptad repeat (NHR) domain in gp41 of the HIV-1 CRF07_BC strain and analyze its molecular structure and antigenicity.</p><p><b>METHODS</b>Overlapping PCR was used to amplify the DNA fragment encoding N51Fd gene, which was then subcloned into the vector pFUSE-hIgG1-Fc2. The construct was confirmed by DNA sequencing. The structure and antigenicity of the recombinant protein N51FdFc-BC were analyzed using bioinformatic software, circular dichroism, and Western blotting.</p><p><b>RESULTS</b>A recombinant expression vector pFUSE/N51Fd-BC was successfully constructed. N51FdFc-BC recombinant protein with a relative molecular mass of about 35 000 was effectively expressed in mammalian 293T cells and could be recognized by rabbit antibodies against HIV-1 gp41 N/C peptides as shown by Western blotting. Bioinformatic analysis showed that the recombinant protein N51FdFc-BC, with a relative molecular mass of 34 315.1 and a PI of 7.59, formed a secondary structure of random coil to allow its interactions as an antigen with antibodies. Circular dichroism measurement confirmed the random coil structure of N51FdFc-BC protein.</p><p><b>CONCLUSION</b>The recombinant protein N51FdFc-BC has a random coil structure and can be used as an immunogen for development of HIV-1 subunit vaccine.</p>
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Asunto principal:
Proteínas Recombinantes
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Proteína gp41 de Envoltorio del VIH
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Química
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Secuencia de Aminoácidos
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VIH-1
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Análisis de Secuencia de ADN
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Estructura Secundaria de Proteína
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Alergia e Inmunología
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Células HEK293
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Genética
Límite:
Humans
Idioma:
Zh
Revista:
Journal of Southern Medical University
Año:
2012
Tipo del documento:
Article