Cloning of the glyceraldehydes 3-phosphate dehydrogenase gene of porphyromonas gingivalis and its expression in E. coli / 华西口腔医学杂志
West China Journal of Stomatology
;
(6): 199-202, 2011.
Artigo
em Chinês
| WPRIM
| ID: wpr-235087
ABSTRACT
<p><b>OBJECTIVE</b>To clone the glyceraldehydes 3-phosphate dehydrogenase (GAPDH) gene of Porphyromonas gingivalis (P. gingivalis) and to induce its fusion expression in E. coli.</p><p><b>METHODS</b>GAPDH was obtained by PCR and was inserted into cloning vector pMD-18-T to construct clone recon. Double enzymes digest the recon pMD18-T-GAPDH and the prokaryotic expression vector pET-32a and then connect to get the expressing recon pET-32a-GAPDH. The recombinant expression plasmid which had been confirmed by enzymes digestion was transformed to E. coli competent cells BL21 and induced the expression of GAPDH with isopropyl beta-D-1-thiogalactopyranoside (IPTG) of different density.</p><p><b>RESULTS</b>DNA sequencing showed that the fragment was 99.802% the same to the sequence published in NCBI. Under the best density, IPTG could be highly expressed.</p><p><b>CONCLUSION</b>The GAPDH had been successfully cloned and expressed in E. coli which gets ready for the following experiment to study the immunity of GAPDH and the homologues antibody preparation.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Oxirredutases
/
Fosfatos
/
Células Cultivadas
/
Reação em Cadeia da Polimerase
/
Clonagem Molecular
/
Porphyromonas gingivalis
/
Clonagem de Organismos
/
Escherichia coli
/
Vetores Genéticos
/
Gliceraldeído
Idioma:
Chinês
Revista:
West China Journal of Stomatology
Ano de publicação:
2011
Tipo de documento:
Artigo
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