Construction of psm-negative mutans of S.mutans UA159
- VernacularTitle:变形链球菌UA159磷酸蔗糖变位酶基因功能丧失菌株的构建
- Author:
Jin DUAN
;
Xiaodi LIU
;
Lihong GUO
- Publication Type:Journal Article
- Keywords:
Streptococcus mutans;
Phospho-sugar mutase gene;
Homologous recombination;
Natural transformation
- From:
Journal of Practical Stomatology
2001;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To knock the phospho-sugar mutase gene (psm) out of the genome of S. mutans strain UA159 and to construct the mutant will lay a foundation for the study of the function of the psm. Methods:Two upstream and downstream DNA sequences of the psm were selected and cloned respectively into multiple cloning sites I and II of suicide plasmid pFW5 to construct the recombinant plasmid which was confirmed by enzyme digesting and sequencing. The recombinant plasmid was transformed into S. mutans UA159 by natural transformation and antibiotic was used to screen the positive transformants. According to the principle of homologous recombination, allelic exchange between the recombinant plasmid and S. mutans UA159 was achieved. Results:By PCR analysis and sequencing, it was confirmed that the psm of S. mutans UA159 was substituted for resistant gene of spectinomycin. Conclusion: The psm-negative mutants of S. mutans UA159 is successfully constructed.
