Detection of quorum-sensing pathway and construction of luxS gene allelic exchange plasmid of Streptococcus mutans
10.3321/j.issn:1002-0098.2008.01.011
- VernacularTitle:变形链球菌AI-2活性测定及luxS基因同源重组质粒的构建
- Author:
Dan-Ni YU
1
;
Yu-Zhi HAN
;
Fu-Sheng HAN
;
Jia CHEN
Author Information
1. 天津医科大学第二医院
- Keywords:
Streptococcus mutans;
Autocrine communication;
Gene;
Recombinated plasmid
- From:
Chinese Journal of Stomatology
2008;43(1):37-40
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect the AI-2 quorum-sensing pathway and construct the luxS g-ene allelic exchange plasmid of Streptococcus mutans. Methods To dectect AI-2 pathway in Streptococcus mutans,the Vibrio harveyi BB170 was used as reporter strain. The PCR fragments of the upstream and downstream regions of luxS and the Erythromycin resistance gene were amplified with the primers respetively,and these fragments were ligated into pUC19 vector with double endonuclease reaction sequentially,the ligated DNAs were transformed into Escherichia coli DH5α,then the reconstructed plasmids were isolated and identified by restricted endonuclease digestions. Results Streptococcus mutans Ingbritt C could induce luminescene of BB170, suggesting the presense of AI-2 quorum sensing pathway in Streptococcus mutans,and such stimulatory activity was maximal at the mid-log growth phase.The recombinant plasmid pUCluxKO was digested by PstI-BamHI,and the digest product were 1000 bp and 5000 bp.When the pUCluxKO Was digested by BamHI-KpnI,the digest product were 1500 bp and 4500 bp.While it was digested by KpnI-ECORI,the digest product were 1000 bp and 5000 bp. All PCR product was in a single belt respectively.Conclusions The recombinant plasmid was cloned effectively and can be used in the construction of S. mutans luxS mutant.
