포도당배지에서 DNA chip을 이용한 자일리톨 감성균주와 내성균주의 유전자 발현 비교
Gene expression profile analysis of xylitol-sensitive and xylitol-resistant Streptococcus mutans in 0.5% glucose containing TYE media using DNA chip
임상욱, 안상헌, 박동옥, 최연희, 송근배,
소속 상세정보
임상욱 ( Im Sang-Uk ) - 경북대학교 치의학전문대학원 예방치과학교실
안상헌 ( Ahn Sang-Hun ) - 대구보건대학 치위생과
박동옥 ( Park Dong-Ok ) - 대구보건대학 치위생과
최연희 ( Choi Youn-Hee ) - 경북대학교 치의학전문대학원 예방치과학교실
송근배 ( Song Keun-Bae ) - 경북대학교 치의학전문대학원 예방치과학교실
KMID : 0355420120360040251
Abstract
Objectives: Streptococcus mutans (S. mutans) is the major causative bacteria in dental caries. Xylitol is an effective anticarious natural sugar substitute by inhibiting the virulence of S. mutans. However, longterm xylitol consumption leads to the emergence of the xylitol-resistant S. mutans (XR). The aim of this study is to analyze the difference of gene expression profile of xylitol-sensitive S. mutans (XS) and XR in 0.5% glucose containing TYE media, using a DNA chip.
Methods: S. mutans KCTC3065 was maintained in 0.5% glucose and 1% xylitol containing TYE media, during 30 days at 37oC 10% CO2 to form XR. The same procedures without xylitol were repeated for the formation of XS. Both XS and XR were cultured in 0.5% glucose with or without 1% xylitol containing TYE media overnight and total RNA was extracted. RNA from XS was labeled with Cy-3 dye as control, and XR were labeled with Cy-5 as references. DNA chip was hybridized for 18-20 h at 42oC.
Results: A total of 277 genes of DNA chip data were significantly increased or decreased in XR. There is a total of 174 XR up-regulated genes in 0.5% glucose and 1% xylitol containing TYE media, and a total of 103 down-regulated genes. For compare with results of DNA chip, 11 in up-regulated genes and 10 in down-regulated were verified by RT-PCR. The most abundant increased genes in XR were related to cell envelope, cellular processes, DNA metabolism, transcription, and protein folding and stabilization. The decreased genes in XR were related to amino acid biosynthesis, toxin production and resistance, energy metabolism, ribosomal proteins synthesis, and signal transduction.
Conclusions: These results indicate that the difference of gene expression profile of XS and XR may be in existence. In particular, results of this study for XR up-regulated genes have a lot of similarities with the already published xylitol-related researches and other functional studies.
키워드
DNA chip;Streptococcus mutans;Virulence;XR;XS;Xylitol
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