Construction of recombinant plasmid with Porphyromonas gingivalis FimA deficiency.
- Author:
Jie YANG
1
;
Kuan-Yu LI
;
Yu LIU
;
Juan WU
;
Wei-Bin SUN
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; DNA, Bacterial; genetics; Fimbriae Proteins; genetics; Gene Knockout Techniques; Genes, Bacterial; Genetic Vectors; Plasmids; genetics; Porphyromonas gingivalis; genetics; Recombinant Proteins; genetics; Sequence Analysis, DNA
- From: Chinese Journal of Stomatology 2012;47(11):671-674
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the recombinant plasmid pPHU281_A_Spec_B, which knock out Porphyrmonas gingivalis (Pg) FimA gene.
METHODSGenomic DNA was extracted from PgATCC33277 which was cultured in anaerobic condition. The upstream and downstream gene of FimA was cloned from Pg genenomic DNA with specific restriction sites by polymerase chain reaction. Suicide vector pPHU281 was inserted by three fragments: upstream, downstream of FimA gene and spectinomycin resistance gene. The recombinant plasmid was confirmed by electrophoresis and sequenced after amplification in compentent cells DH-5α.
RESULTSThe gene sequence was identified by DNA sequencing analysis. The recombinant plasmid pPHU281_A_Spec_B was successfully constructed.
CONCLUSIONSThe recombinant plasmid pPHU281_A_Spec_B was constructed, which may be used for the constructon of FimA deficient Pg.
