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
Dans Chinois
| WPRIM
| ID: wpr-352344
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>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Protéines recombinantes
/
Protéine d'enveloppe gp41 du VIH
/
Chimie
/
Séquence d'acides aminés
/
VIH-1 (Virus de l'Immunodéficience Humaine de type 1)
/
Analyse de séquence d'ADN
/
Structure secondaire des protéines
/
Allergie et immunologie
/
Cellules HEK293
/
Génétique
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2012
Type:
Article
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