1.Genetic diversity of F-ATPase subunits gene uncA amplified from Streptococcus mutans clinical isolates.
De-qin YANG ; Tian-jia LIU ; Heng ZHUANG ; Qing-guo QI ; Song LI ; Jian-guo LIU
Chinese Journal of Stomatology 2006;41(1):29-32
OBJECTIVETo study the genetic diversity of F-ATPase alpha subunit gene uncA derived from Streptococcus mutans (S. mutans) clinical isolates and to investigate the relationship between the genetic diversity of acidurance factor and S. mutans aciduric ability, also and the cariogenicity.
METHODSSixty-four S. mutans strains derived from 34 caries-active individuals and 30 caries-free individuals, including 18 strains displaying high acid tolerance and 20 strains displaying low acid tolerance. Gene uncA was amplified with specific primers from S. mutans genomic DNA, then the PCR products were analyzed by RFLP and sequenced.
RESULTSTwo genotypes A and B of PCR-RFLP were revealed when digested with Hph I. Mbo II also produced two different pattern C and D. The distributions of A and B genotype strains with different caries-sensitivity groups were different (P < 0.05), and the proportion of A genotype strains from caries-activity group was higher than that from caries-free one. The distributions of C and D genotype strains with different acidurance strains were different (P < 0.05), and the proportion of C genotype strains from high acid tolerance group was higher than that from low acid tolerance group. These amplified uncA genes from different group were sequenced and there existed variation of Hph I and Mbo II recognized sites.
CONCLUSIONSThis study indicates that uncA gene of S. mutans F-ATPase obviously displayed genetic diversity. The different Hph I-RFLP and Mbo II-RFLP genotypes could be related to the cariogenicity and acid tolerance of S. mutans strains.
Bacterial Proton-Translocating ATPases ; genetics ; Dental Caries ; microbiology ; Genes, Bacterial ; Genotype ; Humans ; Polymorphism, Restriction Fragment Length ; Streptococcus mutans ; enzymology ; genetics
2.Genetic diversity of ATP synthase cab subunits amplified from Streptococcus mutans clinical isolates from Uyghur children with different caries susceptibility.
Zhen-hua LIU ; Bing-jie LIAN ; Jin ZHAO
Chinese Journal of Stomatology 2012;47(9):547-551
OBJECTIVETo investigate the aciduricity and genetic diversity of ATP synthase subunit gene uncEBF derived from Uyghur children Streptococcus mutans (Sm) clinical isolates and the relationship between the genetic diversity of ATP synthase and Sm aciduric ability and caries susceptibility.
METHODSForty-one Sm strains derived from 24 caries-active individuals and 17 caries-free individuals, including 16 strains displaying high acid tolerance and 17 strains displaying low acid tolerance. Solutions of all isolated Sm with same density were made and cultured at pH 4.0 to 7.0 brain heart infusion (BHI) liquid. Terminal growth situation was compared. Gene uncEBF of these isolates were amplified with specific primers from Sm genomic DNA, and the polymerase chain reaction (PCR) products were analyzed by PCR-restriction fragment length polymorphism (RFLP) and sequenced.
RESULTSAciduric ability of Sm isolated from the high caries-susceptible children were higher than that isolated from caries-free group (P = 0.023). Alu I digested fragments of uncEBF displayed two different patterns A and B. The distributions of A and B genotype strains with different acidurance were different (P = 0.039). A genotype included 7 strains displaying high acid tolerance and 2 strains displaying low acid tolerance;B genotype included 9 strains displaying high acid tolerance and 15 strains displaying low acid tolerance. The distributions of A and B genotype strains in different caries-sensitivity groups were different (P = 0.009). A genotype included 7 high caries-susceptible strains and 12 caries-free strains; B genotype included 17 high caries-susceptible strains and 5 caries-free strain. Some of these amplified uncEBF genes from different genotype were sequenced and testified that there existed variation of Alu I recognized sites.
CONCLUSIONSThe high cariogenecity of Sm strains isolated from caries-active children shows a close relationship with the high aciduric ability of the isolated Sm strains. uncEBF gene of Sm F-ATPase obviously exhibits genetic diversity.
Bacterial Proton-Translocating ATPases ; genetics ; metabolism ; Child, Preschool ; China ; ethnology ; Dental Caries ; microbiology ; Dental Caries Susceptibility ; Genetic Variation ; Genotype ; Humans ; Hydrogen-Ion Concentration ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Streptococcus mutans ; enzymology ; isolation & purification