[Construction of the hemagglutinin-2-deficient mutant of Porphyromonas gingivalis].
OBJECTIVE
To construct and identify the Porphyromonas gingivalis(Pg)ATCC33277 hemagglutinin-2(HA-2)-deficient mutant.


METHODS
The genomic DNA of Pg was isolated from PgATCC33277. The up/down stream genes of HA-2-HA(u), HA(l) were amplified by PCR, and inserted into pSY118 separately which contains a 2.1 kb antibiotic resistance ermF-ermAM cassette. The resultant recombinant plasmid-pSY118-HA was linearized as the gene targating fragment HA-ermF-ermAM and used in the electroporation of PgATCC33277. The Pg HA-2-deficient mutant was screened by allelic exchange. The test of aggregation of red blood cells was used to investigate the function change between PgHA2-deficient mutant and the wild type of PgATCC33277.


RESULTS
The PgHA-2-deficient mutant was identified by PCR. The ability of Pg HA-2-deficient mutant to aggregate red blood cell was significantly decreased compared with the wild type.


CONCLUSIONS
HA-2-deficient mutant of Pg ATCC33277 was constructed successfully, which lays a foundation for further study of its biological function.
Lin
Mei
M
Beijing Institute for Dental Research, Capital Medical University School of Stomatology, Beijing 100050, China.
Yang
Qiu-bo
QB
Yang
Sheng-hui
SH
D000483:Alleles; D004903:Erythrocyte Aggregation; D006388:Hemagglutinins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis / Q000235:genetics*
D006388:Hemagglutinins