Construction of the hemagglutinin-2-deficient mutant of Porphyromonas gingivalis

VernacularTitle:牙龈卟啉单胞菌血凝素2基因缺陷型突变株的构建
Author: Mei LIN ¹ ; Qiu-Bo YANG ; Sheng-Hui YANG
Author Information

1. 首都医科大学附属北京朝阳医院
Keywords: Porphyromonas gingivalis; Hemagglutinins; Gene targeting; Blood coagulation
From: Chinese Journal of Stomatology 2009;44(11):672-676
CountryChina
Language:Chinese
Abstract: Objective To construct and identify the Porphyromonas gingivalis(Pg)ATCC33277hemagglutinin-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_1 were amplified by PCR,and inserted into pSY118 separately which contains a 2.1 kb antibiotic resistance ermF-ermAM cassette.The resultant 0recombinant 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.Condusions HA-2-deficient mutant of Pg ATCC33277 was constructed successfully,which lays a foundation for further study of its biological function.