Design of signal peptide bombyxin and its effect on secretory expression efficiency and levels of Helicobacter pylori urease subunit B in silkworm cells and larvae
Título del documento: Design of signal peptide bombyxin and its effect on secretory expression efficiency and levels of Helicobacter pylori urease subunit B in silkworm cells and larvae
Revue: Brazilian archives of biology and technology
Base de datos: PERIÓDICA
Número de sistema: 000387668
ISSN: 1516-8913
Autores: 1
2
1
1
1
1
1
1
Instituciones: 1Anhui Science and Technology University, Fengyang, Anhui. China
2University of Science and Technology of China, School of Life Science, Hefei, Anhui. China
Año:
Periodo: May-Jun
Volumen: 58
Número: 3
Paginación: 319-325
País: Brasil
Idioma: Inglés
Tipo de documento: Artículo
Enfoque: Experimental, aplicado
Resumen en inglés This study employed a Bac-to-Bac/Bombyx mori bioreactor to mass-produce immunogenic urease subunit B (UreB) from Helicobacter pylori. The signal peptide bombyxin from B. mori was used to promote secretory expression to improve expression levels and was designed and integrated into the UreB gene to generate the Bacmid/BmNPV/(signal peptide)-UreB baculovirus expression system. To determine whether the bombyxin signal peptide resulted in secretory expression of recombinant UreB (rUreB) and to determine the secretory efficiency, we tested the secretory expression level of rUreB in Bm5 cells using ELISA. To further investigate whether secretory expression affected cell viability, cells were evaluated using 0.4% trypan blue staining, and Bacmid/BmNPV/UreB without the signal peptide served as a control. The above recombinant bacmid constructs were injected to silkworm larvae, and the secretory expression level of rUreB was detected using SDS-PAGE and semi-quantitative western blot analysis. The results indicated that the bombyxin signal peptide directed the secretory expression of rUreB and that this expression improved the viability of Bm5 cells. Moreover, the results showed that the expression level of rUreB was 1.5 times higher with the Bacmid/BmNPV constructs containing the bombyxin signal sequence than those without the signal sequence. These results demonstrate that secretory expression can enhance rUreB expression levels and is likely to aid in the large-scale expression and yield of rUreB in silkworm larvae
Disciplinas: Biología,
Química
Palabras clave: Genética,
Proteínas recombinantes,
Expresión génica,
Baculovirus,
Ureasa,
Helicobacter pylori,
Bombyx mori
Keyword: Biology,
Chemistry,
Genetics,
Recombinant proteins,
Gene expression,
Baculovirus,
Urease,
Helicobacter pylori,
Bombyx mori
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