Study On The Relationship Between Chloride Channel Protein Ericf1/ericf2 And Enhanced Fluoride Resistance Of Streptococcus Mutans And Fluoride-resistant Strains

Objective : Through transcriptome sequencing and gene function analysis of Streptococcus mutans?UA-159?and Streptococcus mutans fluoride-resistant strain?UA-159FR?,the biological information and regulatory mechanism were revealed,and the chloride channel protein ericf1 / ericf2 was revealed in different Under the conditions of fluorine resistance,a reasonable guess can be made based on the expected transcriptional regulation mechanism of the protein.In order to explore the mechanism of fluoride resistance of genes related to fluoride resistance of S.mutans strains.Methods: In this experiment,by the cultivation of Streptococcus mutans and fluoride-resistant strains,high-throughput Illumina Hiseq TM2500 sequencing technology was used to sequence the transcriptome of Streptococcus mutans and fluoride-resistant strains,and all gene expression differences were found.Aiming at the difference in expression of genes encoding the corresponding proteins of streptococcus mutans and fluoride-resistant strains of ericf1 / ericf2,fluorescence quantitative polymerase chain reaction?q-PCR?method was used to detect the difference in the expression level of m RNA encoding ericf1 / ericf2 in the two strains.Results: Compared to their parental DEGs?differential-expressed genes?,Streptococcus mutans has 765 up-regulated differential genes and 829 down-regulated differential genes.By comparing the KEGG enrichment pathway with BLAST,61 metabolic pathways were enriched by the KEGG pathway entry,of which 1594 were differential genes.Among them,the ericf1 / ericf2 encoding genes?SMU₁289C,SMU₁290C?encoding the fluoride ion transporter proteins?SMU₁289C,SMU₁290C?,which are involved in fluoride-resistant strains of Streptococcus mutans,were significantly up-regulated compared with the parent strains,and DEGs were significantly different.The expression levels of FPKM encoding genes SMU₁289C and SMU₁290C of Streptococcus mutans fluorine-resistant strains were significantly higher than the parent strains?p <0.05?.Through q-PCR verification,it was found that fluoride-resistant strains were more resistant to fluoride strains than ericf1,the relative expression of ericf2 gene was significantly increased;the relative expression of ericf1 / ericf2 gene was significantly lower in the 1g / LNa F environment than in the fluoride-free environment.Consistent with the results of transcriptome sequencing showing that the fluorine-resistant group was significantly up-regulated compared to the parent group,the results of transcriptome sequencing were further verified.Conclusion: When RNA-seq technology is used to detect the transcriptome of Streptococcus mutans and its fluorine-resistant strains,the sequencing depth,coverage and genome comparison results are ideal,which can basically reflect the overall transcriptome of Streptococcus mutans and its fluorine-resistant strain,Phred numerical detection of each base sequencing error rate in each group is within the ideal range.It provides a platform for revealing differential gene expression of Streptococcus mutans and fluoride-resistant strains,and provides a reference basis for further research on the fluorine resistance mechanism of Streptococcus mutans.According to the KEGG enrichment analysis of differentially expressed genes?DEGs?,it is known that gene information is comprehensively obtained through transcriptome sequencing and bioinformatics analysis,and differential genes are mined from big data.Studies show that F-transportation-related ericf1,the genes encoding ericf2 were significantly different in Pathway enrichment analysis and GO functional enrichment analysis,indicating that they play an important role in the fluorine resistance mechanism of Streptococcus mutans.In PCR verification,the same results were obtained as the transcriptome.These results indicate that the ericf1 and ericf2 related genes in S.mutans are resistant to fluoride.This experiment provides a theoretical basis for studying the prevention of dental caries and the study of fluoride resistance of fluoride-resistant strains.It provides favorable information for the study of the fluorine resistance mechanism of Streptococcus mutans and provides a good reference for the subsequent proteome research.