Construction and cellular expression of GTF-PAc fusion anti-caries DNA vaccine / 中华口腔医学杂志
Chinese Journal of Stomatology
; (12): 456-458, 2002.
Article
en Zh
| WPRIM
| ID: wpr-347327
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct a fusion anti-caries DNA vaccine pGLUA-P carrying GLU fragment from gtfB gene of Streptococcus mutans GS-5 and A-P fragment including the A region and P region of PAc protein from a DNA anti-caries vaccine pCIA-P, and to investigate its expression in prokaryotic and eukaryotic cells.</p><p><b>METHODS</b>The sequence of GLU fragment in pGLU plasmid was testified by DNA sequencing. The fusion anti-caries DNA vaccine was constructed by ligating A-P fragment from pCIA-P to pGLU. The expression of GLUA-P fusion protein in E. coli BL21 (DE3) was induced by IPTG and checked by SDS-PAGE electrophoresis. pGLUA-P was transfected in vitro to cultured rat primary muscle cells by cation liposome Dosper, and immunohistochemical method was used to test the expression of GLUA-P fusion protein in cells.</p><p><b>RESULTS</b>GLU sequence was identical with relative sequence of GTF-I (GS-5 strain) in Gene Bank. Recombinant eukaryotic expression plasmid pGLUA-P was confirmed to have both GLU and A-P fragment. After pGLUA-P was transferred into E. coli (DE3), it could express a new 115 000 protein by the induce of IPTG. Specific brown products could be found in the cytoplasm of cultured rat primary muscle cells transfected by pGLUA-P.</p><p><b>CONCLUSIONS</b>Fusion anti-caries DNA vaccine pGLUA-P is successfully constructed and confirmed by sequencing and enzymes digestion. Fusion GLUA-P protein can be correctly expressed in prokaryotic and eukaryotic cells.</p>
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Plásmidos
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Streptococcus mutans
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Proteínas Bacterianas
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Proteínas Recombinantes de Fusión
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Glicoproteínas de Membrana
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Transfección
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Expresión Génica
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Células Cultivadas
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Clonación Molecular
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Ratas Wistar
Límite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Stomatology
Año:
2002
Tipo del documento:
Article