The Influence Of SiRNA Interfered Ffh, HtrA Gene On Virulence Factors Of Fluoride-resistant Of Streptococcus Mutans

Purpose:Caries is a very common oral diseases, is caused by various factors working togetherdental hard tissue caused by loss of sexually transmitted diseases. Wherein the bacterialinfection is an important factor in the occurrence of dental caries[1.2.3], in particular, moreoften in children[4]. In the study of microbiological characteristics of caries showedStreptococcus mutans[5]is the strongest caries cariogenic bacteria[6-7]. Acid and acidproduction is an important factor of S. mutans in the oral environment, especially on thesurvival of dental biofilms[8]. Study[3]showed that: closely related to the production ofacid, acid and biofilm formation of Streptococcus mutans ffh expression htrA geneexpression and gene. In this study by comparing the siRNA[9]targeted gene silencing ffhrespectively[10-11]and htrA[12]After the gene of Streptococcus mutans in strong acid weakacid and the role of various sugars to study the production of acid fluoride-resistant strains,acid impact resistance and biological characteristics of biofilm formation[13]and so on.Reveals a deeper Streptococcus mutans acid mechanism, but also provide new ideas andtheoretical guidance for the prevention and treatment of dental caries.Method:Vitro fluoride-resistant Streptococcus mutans bacteria in accordance with theconventional method in the experimental group after the identification of Streptococcusmutans to50ug/ml NaF into a sliding scale until the fluoride-resistant colonies there1000ug/ml NaF of BHI solid medium.Electroporation transfection identified after UA159-FR suspension with siRNA oligo2:1mixing ratio joined electroporation cup electrode choose electrodes. With real time-PCRmethod to detect and filter out the effect of transfection of siRNA sequences with betterconditions.Function test of the transfected cultured UA159-FR0.5intervals, detecting thecollected configuration data the acid pH for from3.5to7.0of the culture medium andanalyzed; be produced in the culture medium containing5%BHI different saccharidedetecting the collected data and analyzed acid; biofilm formation was observed in cultureplaced coverslip[14]the plates.Results: siRNA oligo in fluoride-resistant strains of Streptococcus mutans ffh successfulinterference and htrA gene expression. Application of real-time PCR technology for foursiRNA sequences were screened group obtained better interference siRNA sequences forsubsequent experiments.Interference after UA159-FR be produced acid detection, interference data htrA gene issuperior interference ffh data genes. SiRNA interference with Streptococcus mutans provedto be effective. Detection results were defined as24hours of time in order to compare dataand analysis.Biofilm build process, the interference htrA gene UA159-FR can be observed notcomplete biofilm similar results; interference ffh gene UA159-FR biofilm structureobserved similar, but with a pore size ranging from the structure.Conclusion:SiRNA sequences electroporation successfully transfected into fluoride-resistant strainsof Streptococcus mutans were successful gene expression were interfering fluoride-resistantstrains of Streptococcus mutans ffh and htrA gene, and successfully detected the interferencefunction, acid-resistant and acid-derived biofilm samples and related test data and analysis.