Construction PG0839 gene-defective mutant of Porphyromonas gingivalis / 华西口腔医学杂志
West China Journal of Stomatology
; (6): 192-200, 2012.
Article
en Zh
| WPRIM
| ID: wpr-241830
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>In order to determine the function of PG0839 gene from Porphyromonas gingivalis (P. gingivalis) W83 strains, we intended to create a mutant in the PG0839 gene by homologous recombination.</p><p><b>METHODS</b>1 584bp PG0839 gene fragment was amplified, digested by BamH I and EcoR I, purified and ligated to pUC19. The recombinant plasmid was designated as pPG0839-1. The erm cassette (2 101 bp) was inserted into the EcoR V restriction site of the PG0839 gene. The resultant recombinant plasmid, pPG0839-2, was used as a donor in the electroporation of P. gingivalis W83. After electroporated and selected on erythromycin brain heart infusion plates, a single colony was collected and designated as PG0839 gene-defective mutant.</p><p><b>RESULTS</b>A mutant in PG0839 gene was created by insertional inactivation, and inactivation of PG0839 gene was confirmed by restriction endonuclease digestive, sequencing, polymerase chain reaction (PCR) and reverse transcription PCR.</p><p><b>CONCLUSION</b>A PG0839 gene-defective mutant was created successfully.</p>
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WPRIM
Asunto principal:
Secuencia de Bases
/
Reacción en Cadena de la Polimerasa
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Porphyromonas gingivalis
/
Genes Bacterianos
Idioma:
Zh
Revista:
West China Journal of Stomatology
Año:
2012
Tipo del documento:
Article