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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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Generation of antibodies against N-terminus fragment of AgI/II protein from Streptococcus mutans GS-5 (연쇄상구균(Streptococcus mutans GS-5)의 항원단백질 AgI/II의 N-terminus절편에 대한 항체형성)

  • Han, Ji-Hye;Baik, Byeong-Ju;Yang, Yeon-Mi;Park, Jeong-Yeol;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.3
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    • pp.401-410
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    • 2006
  • Dental caries results from localized demineralization of tooth enamel by acids of bacterial origin produced from the fermentation of dietary sugars. A group of related oral bacteria, collectively known as mutans streptococci, are implicated as the primary etiological agents of human caries. Within this group, Streptococcus mutans has been known as 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 their virulence Cell-surface fibrillar proteins, which mediate adherence to the salivary pellicle are virulence components of mutans streptococci, and primary candidates for a human caries vaccine. Here we report that the AgI/II gene from S. mutans GS-5 were cloned by PCR amplification of the bacterial chromosomal DNA and the integrity of cloned genes were confirmed by nucleotide sequencing. Sequence analyses showed the sequence alignment of 280 nucleotides between the cloned AgI/II and the reported sequence of S. mutans GS-5 showed the perfect match The cloned genes which signal nucleotide was truncated, were transferred into bacterial expression vector and the recombinant proteins were purified as His-tag fusion proteins In order to generate polyclonal antibodies against the recombinant proteins, AgI/II mr, some $100{\mu}g$ of the proteins was injected into mice three times. It can be used for an effective vaccine production to prevent dental caries caused by pathogenic S. mutans.

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Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria×ananassa Duchesne) (딸기 흰가루병 저항성 계통 선발을 위한 분자마커 개발)

  • Je, Hee-Jeong;Ahn, Jae-Wook;Yoon, Hae-Suk;Kim, Min-Keun;Ryu, Jae-San;Hong, Kwang-Pyo;Lee, Sang-Dae;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.722-729
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    • 2015
  • Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

Analysis of Spatial Trip Regularity using Trajectory Data in Urban Areas (도시부 경로자료를 이용한 통행의 공간적 규칙성 분석)

  • Lee, Su jin;Jang, Ki tae
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.17 no.6
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    • pp.96-110
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    • 2018
  • As the development of ICT has made it easier to collect various traffic information, research on creating new traffic attributes is drawing attention. Estimation and forecasts of demand and traffic volume are one of the main indicators that are essential to traffic operation, assuming that the traffic pattern at a particular node or link is repeated. Traditionally, a survey method was used to demonstrate this similarity on trip behavior. However, the method was limited to achieving high accuracy with high costs and responses that relied on the respondents' memory. Recently, as traffic data has become easier to gather through ETC system, smart card, studies are performed to identify the regularity of trip in various ways. In, this study, route-level trip data collected in Daegu metropolitan city were analyzed to confirm that individual traveler forms a spatially similar trip chain over several days. For this purpose, we newly define the concept of spatial trip regularity and assess the spatial difference between daily trip chains using the sequence alignment algorithm, Dynamic Time Warping. In addition, we will discuss the applications as the indicators of fixed traffic demand and transportation services.

PCR-based markers for discriminating Solanum demissum were developed by comparison of complete chloroplast genome sequences of Solanum species (가지속 식물의 엽록체 전장유전체 비교를 통한 PCR 기반의 Solanum demissum 특이적 분자마커 개발)

  • Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.18-25
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    • 2021
  • Solanum demissum is one of the wild Solanum species originating from Mexico. It has wildly been used for potato breeding due to its resistance to Phytophthora infestans. S. demissum has an EBN value of four, which is same as that of S. tuberosum, so that it is directly crossable for breeding purposes with the cultivated tetraploid potato (S. tuberosum). In this study, the chloroplast genome sequence of S. demissum obtained by next-generation sequencing technology was described and compared with those of seven other Solanum species to develop S. demissum-specific markers. Thetotal sequence length of the chloroplast genome is 155,558 bp, and its structural organization is similar to those of other Solanum species. Phylogenetic analysis with ten other Solanaceae species revealed that S. demissum is most closely grouped with S. hougasii and S. stoloniferum followed by S. berthaultii and S. tuberosum. Additional comparison of the chloroplast genome sequence with those of seven other Solanum species revealed two InDels specific to S. demissum. Based on these InDels, two PCR-based markers for discriminating S. demissum from other Solanum species were developed. The results obtained in this study will provide an opportunity to investigate more detailed evolutionary and breeding aspects in Solanum species.