Abstract
ABSTRACT Acid resistance is one of key properties assisting the survival of cariogenic bacteria in a dental caries environment, but only a few genes conferring acid resistance have been identified to data. Functional metagenomics provides a systematic method for investigating commensal DNA to identify genes that encode target functions. Here, the host strain Escherichia coli DH10B and a constructed bidirectional transcription vector pSKII+-lacZ contributed to the construction of a metagenomic library, and 46.6 Mb of metagenomic DNA was cloned from carious supragingival plaque of 8children along with screening for lethal functionality. The screen identified 2 positive clones that exhibited a similar aciduric phenotype to that of the positive controls. Bioinformatic analysis revealed that these two genes encoded an ATP/GTP-binding protein and a malate dehydrogenase. Moreover, we also performed functional screening of Streptococcus mutans, since it is one of the predominant cariogenic strains but was not identified in our initial screening. Five positive clones were retrieved. In conclusion, our improved functional metagenomics screening method helped in the identification of important acid resistance genes, thereby providing new insights into the mechanism underlying caries formation as well as in the prevention and treatment of early childhood caries (ECC).
Highlights
Childhood caries (ECC) is one of the most common infectious diseases in children and affects up to 60–90% of children worldwide [1]
Acid resistance is a key factor for the survival of cariogenic bacteria [5] in the typical carious microenvironment, in which the pH values fluctuate in the range from pH 7 to 3, primarily due to diet [6]
Analysis found that this could be attributed to the start codon ATG of the vectorself aiding in the generation of a fusion protein weakening the expression of green fluorescent protein (GFP)
Summary
Childhood caries (ECC) is one of the most common infectious diseases in children and affects up to 60–90% of children worldwide [1]. The prevention and treatment of ECC are extremely important and mandate intensive study [4]. It is well known that microorganisms living in the dental plaque biofilm play a vital and direct role in the occurrence of tooth decay [2]. Survival and propagation of microorganisms in the caries microenvironment inducing tooth decay require that they exhibit certain properties, such as adhesion, acidogenicity and acid resistance. Acid resistance is a key factor for the survival of cariogenic bacteria [5] in the typical carious microenvironment, in which the pH values fluctuate in the range from pH 7 to 3, primarily due to diet [6]. The acid resistance of oral microbes is taken into account in the study of ECC prevention and treatment, which offers potential new insights into clinical applications and personalized medicine [7]
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