• Title/Summary/Keyword: gtf

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IDENTIFICATION OF THE AG I/II AND GTFD GENES FROM STREPTOCOCCUS MUTANS GS-5 (연쇄상구균 GS-5의 ag I/II와 gtfD 유전자 클로닝)

  • Jeong, Jin-Woo;Baik, Byeong-Ju;Yang, Yeon-Mi;Seo, Jeong-Ah;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.2
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    • pp.357-369
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    • 2005
  • Streptococci are Gram-positive, facultative anaerobes and have no catalase activities. Among mutans streptococci containing ${\alpha}-type$ hemolytic activity, S. mutans is a causative agent for dental caries. As well as acid production yielding the demineralization of tooth enamel, adherence and colonization of S. mutans to the teeth are also important for its virulence. These early colonization are accomplished by the bacterial fibrillar protein, Antigen I/II (Ag I/II) and glucosyltransferase (GTF). Therefore, Ag I/II and GTF are reasonable targets for the development of vaccine against S. mutans GS-5. The ag I/II and gtfD genes from S. mutans GS-5 were cloned and sequenced. Sequence analyses showed the nucleotides sequence of cloned genes had high homology to the sequences previously reported. The sequence alignment of 280 nucleotides between the cloned Ag I/II and the available sequence of the corresponding S. mutans GS-5 showed the perfect match. Comparing with the sequence of gtfD from S. mutans UA159, the corresponding nucleotide sequence of S. mutans GS-5 showed some mismatches and the mismatches introduced changes in four residues out of 105 amino acids, yielding four missense mutations.

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Analysis of Gene Expression in response to acid stress of Streptococcus mutans Isolated from a Korean Child (한국인 아동으로부터 분리한 Streptococcus mutans 의 산 스트레스에 따른 유전자 발현변화 분석)

  • Kang, Kyung-Hee;Kim, Young-Kwon;Lee, Hyung-Suk;Jin, Ing-Ryol
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.10
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    • pp.2990-2996
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    • 2009
  • S. mutans, one of a major causal agents of dental caries, is component of the dental plaque and produces various organic acids such as lactic acid as the end-product of glycolysis. In this study, we are interested in comparing the gene expression of acid-shocked and control cells of S. mutans isolated from Korean with caries. Expression levels of gtfB, gtfC, gtfD and ftf were analyzed by Real-time PCR, when the cells were grown under 20 mM lactic acid stress in the exponential phase. The data showed reduced expression of these genes. S. mutans is known to have developed a variety of mechanisms to tolerate acid sterss. A more detailed analysis of the functions and interactions of acid stress proteins connecting the growth, stress tolerance, biofilm formation is under way.

Comparison of Cariogenic Characteristics between Fluoride-sensitive and Fluoride-resistant Streptococcus mutans (불소 민감성 Streptococcus mutans와 불소 저항성Streptococcus mutans의 우식원성 특성 비교)

  • Ong, Seung-Hwan;Kim, Jongsoo;Baek, Dong-Heon;Yoo, Seunghoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.4
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    • pp.397-405
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    • 2020
  • The aim of this study is to compare cariogenic characteristics of fluoride-sensitive Streptococcus mutans [fluoride-sensitive (FS) S. mutans ] and fluoride-resistant Streptococcus mutans [fluoride-resistant (FR) S. mutans] in the presence of sucrose, and to evaluate its effect on cariogenic biofilm formation. S. mutans ATCC 25175 was continuously cultured in trypticase soy broth (TSB) containing NaF (70 ppm) for 40 days to generate FR S. mutans. FS and FR S. mutans were inoculated in TSB with or without 2% sucrose, and optical density and pH were measured every hour. An oral biofilm was formed using saliva bacteria and analyzed through confocal laser scanning microscopy and CFU count. Finally, the expression of glucosyltransferases genes of both S. mutans was investigated through RT-PCR. FR S. mutans exhibited slower growth and lower acidogenicity in the presence of sucrose compared to FS S. mutans . Both cariogenic and single species biofilm formation was lower in the presence of FR S. mutans, along with reduced number of bacteria. FR S. mutans showed significantly low levels of gtfB, gtfC, and gtfD expression compared to FS S. mutans . On the basis of results, FR S. mutans may be less virulent in the induction of dental caries.

THE COMPARISON OF STREPTOCOCCUS MUTANS ISOLATED FROM OCCLUSAL SURFACES OF CARIES AND NON-CARIES TEETH (우식치아와 정상치아의 교합면에서 분리한 Streptococcus mutans의 비교)

  • Park, Ho-Won;Jung, Tae-Sung;Jung, Jin;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.1
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    • pp.129-141
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    • 2001
  • When oral microorganisms were sampled from occlusal surfaces of caries and non-caries teeth, $3.43\times10^5$ CFU and $3.47\times10^3$ CFU of bacteria were counted on MSB agar plates, respectively. All the 20 colonies isolated from a caries surface were Streptococcus mutans but, only two of 20 colonies were identified as Streptococcus mutans by API test. S. mutans SM1 from caries tooth and S. mutans SM2 from non-caries tooth showed the same results except for $\alpha-galactosidase$ activity on sugar fermentation tests and biochemical tests. For the bacterial replication, both SM1 and SM2 were actively multiplicated at pH 5.5. And the viability of SM1 was high at 20% of sucrose, while that of SM2 was high at 5% of sucrose in the media. SM1 actively replicated at 16mM of $CaCl_2$, 160mM of KCl, and 6.4mM of $MgCl_2$, and the replication of SM2 was increased at 16mM of $CaCl_2$, 40mM of KCl, 6.4mM of $MgCl_2$. At 1mM of sodium bicarbonate and sodium phosphate, both bacteria were actively multiplicated. SM1 and SM2 were actively replicated at 1mM and 10mM of Tris, respectively. For potassium phosphate buffer, SM1 grew well proportionally to the concentration up to 100mM, while the growth of SM2 were inhibited by the increase of concentration. The 4.6 kb of gtf gene was amplified with a pair of primer, gtfB-F961 and gtfC-R5574 by polymerase chain reaction from the chromosomal DNA of SM1 and SM2. When 4.6kb bands were eluted from gel and were treated with restriction enzyme, EcoR I produced the same RFLP like 0.8kb and 3.8kb of DNA fragments for S. mutans GS-5, SM1 and SM2. By Hind III, the PCR products weren't digested for S. mutans GS-5 and SM1, but 3 fragments such as 2.4kb, 1.8kb and 400bp were examined for SM2. These results indicated the difference between gtf genes of SM1 and SM2. BamH I treatment showed 4 fragments for SM1 and SM2, while the 3 fragments for S. mutans GS-5. The PCR products were not digested by Kpn I, Sma I, Xho I and Pst I.

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THE EFFECTS OF IONS AND BUFFER SOLUTIONS ON THE MRNA EXPRESSION OF gtfD GENE OF Streptococcus mutans (Streptococcus mutans의 gtfD 유전자 발현에 대한 이온 및 완충액의 영향)

  • Kim, Bo-Young;Kim, Shin;Chung, Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.2
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    • pp.314-322
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    • 2004
  • The production of a glucan was affected by the concentration of ions and buffer solutions, and nutrients in an oral cavity. In this study, the effects of ions and buffer solutions on the mRNA expression of gtfD gene in Streptococcus mutans, an important causative agent of dental caries, were investigated by Fluorescent in situ hybridization(FISH). At first, ions and buffer solutions had little effect on the multiplication of Streptococcus mutans. The green fluorescence according to the mRNA expression of gtfD gene was detected in the BHI broth containing 1% sucrose. The intensities of the green fluorescence were strong at 0.25mM of $CaCl_2$. Little fluorescence was detected by the addition of KCl, except far 10mM KCl at which fluorescence intensities were similar to those of the control. Fluorescence intensities were weak at each concentration of $MgCl_2$ when compared to the control. As for buffer solutions, fluorescence intensities were similar to those of the control at each concentration of buffer solutions, except that they were little detected at 100mM of potassium phosphate.

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Antibacterial and Antibiofilm Activities of Diospyros malabarica Stem Extract against Streptococcus mutans (Streptococcus mutans에 대한 인도감나무 줄기 추출물의 항균활성 및 생물막 형성 억제 효과)

  • Kim, Hye Soo;Lee, Sang Woo;Sydara, Kongmany;Cho, Soo Jeong
    • Journal of Life Science
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    • v.29 no.1
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    • pp.90-96
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    • 2019
  • The objective of this study was to evaluate the potential of Diospyros malabarica stem extract, a natural materials, in oral health material. With this aim in mind, thin layer chromatography (TLC), TLC-bioautography, high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and real-time qPCR were performed. The antibacterial activity of D. malabarica stem extract against Streptococcus mutans KCTC3065 was confirmed in an n-hexane fraction with low polarity. The molecular weight of the antibacterial compound was estimated to be 188 by ESI-MS analysis. The inhibitory effects of the extract on biofilm formation and gene expression related to biofilm formation of S. mutans were determined by SEM and real-time PCR analysis. The extract inhibited the formation of S. mutans biofilms at D. malabarica stem extract concentrations of 1 mg/ml, as shown by SEM. The real-time PCR analysis showed that the expression of the gtfC gene, which is associated with biofilm formation, was significantly decreased in a dose-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts, a natural materials, can be used in oral health products to suppress the formation of biofilms by inhibiting tooth adhesion of S. mutans, a causative agent of dental caries.

Folded Architecture for Digital Gammatone Filter Used in Speech Processor of Cochlear Implant

  • Karuppuswamy, Rajalakshmi;Arumugam, Kandaswamy;Swathi, Priya M.
    • ETRI Journal
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    • v.35 no.4
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    • pp.697-705
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    • 2013
  • Emerging trends in the area of digital very large scale integration (VLSI) signal processing can lead to a reduction in the cost of the cochlear implant. Digital signal processing algorithms are repetitively used in speech processors for filtering and encoding operations. The critical paths in these algorithms limit the performance of the speech processors. These algorithms must be transformed to accommodate processors designed to be high speed and have less area and low power. This can be realized by basing the design of the auditory filter banks for the processors on digital VLSI signal processing concepts. By applying a folding algorithm to the second-order digital gammatone filter (GTF), the number of multipliers is reduced from five to one and the number of adders is reduced from three to one, without changing the characteristics of the filter. Folded second-order filter sections are cascaded with three similar structures to realize the eighth-order digital GTF whose response is a close match to the human cochlea response. The silicon area is reduced from twenty to four multipliers and from twelve to four adders by using the folding architecture.

Characterization of Streptococcus mutans Ingbritt Sucrose-glucan Glucosyltransferase and the Inhibition Effect of Chitin Derivatives on its Activity (Streptococcus mutans Ingbritt sucrose-glucan glucosyltransferase 특성과 그 활성에 미치는 키틴 유도체들의 효과)

  • Ju, Wan-Taek;Ji, Myeong-Sim;Park, Ro-Dong
    • Journal of Applied Biological Chemistry
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    • v.55 no.3
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    • pp.173-178
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    • 2012
  • Sucrose-glucan glucosyltransferase (Gtf) is an important enzyme involved in the cavity formation process where insoluble glucan is synthesized. In this study, we purified Gtf from Streptcoccus mutans Ingbritt through ammonium sulfate precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex column chromatographies. A 13-fold of purification was achieved with a total yield of 6.3%. The apparent molecular mass of the enzyme was determined to be 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal pH and temperature were established to be 6.0 and $40^{\circ}C$, respectively. The enzyme activity could be inhibited to 22-59% by 1 mM $Hg^{2+}$, $Cu^{2+}$ and $Al^{3+}$, and to 68% by 1 mM EDTA. It was also inhibited 40% by 2 mM xylitol and 35-45% by 0.05% soluble chitosan, glycol chitosan, and glycol chitin. This is the first report to reveal the inhibition effect of chitin derivatives on Gtf activity, which may be further applicable to develop gargles to overcome cavity.

Cloning and expression of Streptococcus mutans GS-5 glucosyltransferase (Streptococcus mutans GS-5 Glucosyltransferase의 클로닝과 발현)

  • Kim, Su-Kyeong;Kim, Jae-Gon;Baik, Byeong-Ju;Yang, Yeon-Mi;Lee, Kyung-Yeol;Park, Jeong-Yeol
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.1
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    • pp.73-82
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    • 2008
  • Dental caries is an infectious disease caused by mutans streptococci, and is a primary etiologic agent of dental caries in humans. The molecular pathogenesis of mutans streptococcal-associated dental caries occurs in three phases. Firstly, S. mutans attaches to tooth surface via a cell surface adhesion termed antigen I/II. In the second phase, the glucosyltransferase(GTFs) synthesize polymers like glucans in the presence of sucrose. In the third phase, the multivalent glucans interacts with glucan binding proteins (GBPs) and they make dental plaque and accumulation of microorganisms. Many studies and clinical trials have indicated that a mucosal immune response to these antigens(Ag I/II, GTFs, GBPs) of S. mutans can influence the pathogenesis of dental caries. So these antigens can be important vaccine candidates for immunologic intervention against dental caries. In this study, we cloned the genes for GTFb, GTFc, GTFd from S. mutans GS-5 and did the nucleotide sequence analysis. And the recombinant proteins of GTFd and N-terminus of GTFd were expressed. Intact GTF which we get from this experiment can be used for antibody production specific for any GTF activity domain through animal experiment.

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