Citation

Yang J, Li KY, Liu Y, Wu J, Sun WB. Zhonghua Kou Qiang Yi Xue Za Zhi 2012; (12): 671-674.

Copyright

(Copyright © 2012, Chinese Medical Association)

DOI

10.3760/cma.j.issn.1002-0098.2012.11.008

PMID

unavailable

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.

Language: zh