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Construction of hsf1 Knockout-mutant of a Thermotolerant Yeast Strain Saccharomyces cerevisiae KNU5377 (고온내성 연료용 알코올 효모균주 Saccharomyces cerevisiae KNU5377에서 HSF1 유전자의 변이주 구축)

  • Kim Il-Sup;Yun Hae-Sun;Choi Hye-Jin;Sohn Ho-Yong;Yu Choon-Bal;Kim Jong-Guk;Jin Ing-Nyol
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.454-458
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    • 2006
  • HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

Development of an Obesity Management Program for Elementary School Children - Based on Keller's Motivation Theory - (초등학생 비만관리프로그램 개발을 위한 예비연구 - Keller의 동기이론을 중심으로 -)

  • Kim, Min-Jeong;Kim, Hee-Soon;Kim, Sun-Ah;Kim, Duk-Hee
    • Child Health Nursing Research
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    • v.13 no.3
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    • pp.257-264
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    • 2007
  • Purpose: The purpose of this study was to develop an effective obesity management program for elementary school children, based on the motivation theory. Method: This study was a methodological study. A child obesity management program, based on the motivation theory was developed, a web site was made and children with obesity participated in the internet program. After the children finished the program, they evaluated the clinical validity of the program. The clinical validity was tested from Jun 12 to 16, 2006. The participants were 6 students. The evaluation tool was Keller's IMMS(Instructional Material Motivation Survey). Data were evaluated using $means{\pm}SD$ for four major components A,R,C,S (Attention, Relevance, Confidence, Satisfaction), based on ARCS in the motivation theory. Results: The program developed in this study was named 'ARCS children obesity escape', and the URL is 'http://www.119kid.co.kr'. Regarding the four components of ARCS, the overall reaction of participants showed that they were very highly motivated by this program. Conclusion: Utilizing the ARCS motivation strategies for solving obesity problems for children is an effective method to motivate the management of obesity, and can increase achievement motivation, attention, relevance, confidence, and satisfaction with the management of obesity. Thus, the program developed in this study is expected to reduce obesity in children, and become an important guide for obesity management and health improvement for children.

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An Integrated Genomic Resource Based on Korean Cattle (Hanwoo) Transcripts

  • Lim, Da-Jeong;Cho, Yong-Min;Lee, Seung-Hwan;Sung, Sam-Sun;Nam, Jung-Rye;Yoon, Du-Hak;Shin, Youn-Hee;Park, Hye-Sun;Kim, Hee-Bal
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.11
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    • pp.1399-1404
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    • 2010
  • We have created a Bovine Genome Database, an integrated genomic resource for Bos taurus, by merging bovine data from various databases and our own data. We produced 55,213 Korean cattle (Hanwoo) ESTs from cDNA libraries from three tissues. We concentrated on genomic information based on Hanwoo transcripts and provided user-friendly search interfaces within the Bovine Genome Database. The genome browser supported alignment results for the various types of data: Hanwoo EST, consensus sequence, human gene, and predicted bovine genes. The database also provides transcript data information, gene annotation, genomic location, sequence and tissue distribution. Users can also explore bovine disease genes based on comparative mapping of homologous genes and can conduct searches centered on genes within user-selected quantitative trait loci (QTL) regions. The Bovine Genome Database can be accessed at http://bgd.nabc.go.kr.

Delineating Transcription Factor Networks Governing Virulence of a Global Human Meningitis Fungal Pathogen, Cryptococcus neoformans

  • Jung, Kwang-Woo;Yang, Dong-Hoon;Maeng, Shinae;Lee, Kyung-Tae;So, Yee-Seul;Hong, Joohyeon;Choi, Jaeyoung;Byun, Hyo-Jeong;Kim, Hyelim;Bang, Soohyun;Song, Min-Hee;Lee, Jang-Won;Kim, Min Su;Kim, Seo-Young;Ji, Je-Hyun;Park, Goun;Kwon, Hyojeong;Cha, Sooyeon;Meyers, Gena Lee;Wang, Li Li;Jang, Jooyoung;Janbon, Guilhem;Adedoyin, Gloria;Kim, Taeyup;Averette, Anna K.;Heitman, Joseph;Cheong, Eunji;Lee, Yong-Hwan;Lee, Yin-Won;Bahn, Yong-Sun
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.59-59
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    • 2015
  • Cryptococcus neoformans causes life-threatening meningoencephalitis in humans, but the treatment of cryptococcosis remains challenging. To develop novel therapeutic targets and approaches, signaling cascades controlling pathogenicity of C. neoformans have been extensively studied but the underlying biological regulatory circuits remain elusive, particularly due to the presence of an evolutionarily divergent set of transcription factors (TFs) in this basidiomycetous fungus. In this study, we constructed a high-quality of 322 signature-tagged gene deletion strains for 155 putative TF genes, which were previously predicted using the DNA-binding domain TF database (http://www.transcriptionfactor.org/). We tested in vivo and in vitro phenotypic traits under 32 distinct growth conditions using 322 TF gene deletion strains. At least one phenotypic trait was exhibited by 145 out of 155 TF mutants (93%) and approximately 85% of the TFs (132/155) have been functionally characterized for the first time in this study. Through high-coverage phenome analysis, we discovered myriad novel TFs that play critical roles in growth, differentiation, virulence-factor (melanin, capsule, and urease) formation, stress responses, antifungal drug resistance, and virulence. Large-scale virulence and infectivity assays in insect (Galleria mellonella) and mouse host models identified 34 novel TFs that are critical for pathogenicity. The genotypic and phenotypic data for each TF are available in the C. neoformans TF phenome database (http://tf.cryptococcus.org). In conclusion, our phenome-based functional analysis of the C. neoformans TF mutant library provides key insights into transcriptional networks of basidiomycetous fungi and ubiquitous human fungal pathogens.

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Linkage Map and Quantitative Trait Loci(QTL) on Pig Chromosome 6 (돼지 염색체 6번의 연관지도 및 양적형질 유전자좌위 탐색)

  • Lee, H.Y.;Choi, B.H.;Kim, T.H.;Park, E.W.;Yoon, D.H.;Lee, H.K.;Jeon, G.J.;Cheong, I.C.;Hong, K.C.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.939-948
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    • 2003
  • The objective of this study was to identify the quantitative traits loci(QTL) for economically important traits such as growth, carcass and meat quality on pig chromosome 6. A three generation resource population was constructed from cross between Korean native boars and Landrace sows. A total of 240 F$_2$ animals were produced using intercross between 10 boars and 31 sows of F$_1$ animals. Phenotypic data including body weight at 3 weeks, backfat thickness, muscle pH, shear force and crude protein level were collected from F$_2$ animals. Animals including grandparents(F$_0$), parents(F$_1$) and offspring(F$_2$) were genotyped for 29 microsatellite markers and PCR-RFLP marker on chromosome 6. The linkage analysis was performed using CRI-MAP software version 2.4(Green et al., 1990) with FIXED option to obtain the map distances. The total length of SSC6 linkage map estimated in this study was 169.3cM. The average distance between adjacent markers was 6.05cM. For mapping of QTL, we used F$_2$ QTL Analysis Servlet of QTL express, a web-based QTL mapping tool(http://qtl.cap.ed.ac.uk). Five QTLs were detected at 5% chromosome-wide level for body weight of 3 weeks of age, shear force, meat pH at 24 hours after slaughtering, backfat thickness and crude protein level on SSC6.

The Web Application of Integrated Sasang Constitutional Diagnosis β-version (통합 체질진단 툴 β-version의 웹기반 응용프로그램)

  • Jin, Hee-Jeong;Kim, Jang-Woong;Kim, Young-Su;Lee, Si-Woo;Jang, Eun-Su
    • Journal of Sasang Constitutional Medicine
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    • v.24 no.1
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    • pp.13-20
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    • 2012
  • 1. Objectives : It is very important to classify people into Sasang constitution correctly in SCM. There have been many researches for this and several tools have been developed for diagnosis of Sasang constitution. In our study, we introduce a new web application for Integrated Sasang Constitutional Diagnosis (ISCD) ${\beta}$-version and algorithm on the base of face, body shape, voice and questionnaire. 2. Development : The web application of ISCD ${\beta}$-version was designed to be used easily for subject, staffs, and oriental medical doctors. For this purpose, we developed a web-application of Integrated Sasang Constitutional Diagnosis ${\beta}$-version using mysql database, tomcat web system, JSP, JAVA, and C++ languages. 3. Current State : The ISCD ${\beta}$-version could be accessed at http://210.218.196.115/SDT/login.jsp. The ISCD ${\beta}$-version consisted of 3 parts, for staffs, subject and oriental medical doctors. The system has been managed since February 2011. Currently 7 oriental hospitals have used the system and 1,439 subjects have been diagnosed by the system. 4. Conclusion and future work : Although many researchers have tried to develop a system or an algorithm for diagnosis of subject's constitution, we could have not used the system based on objective information of human body type, characters, symptoms. In this study, we describe a web application of objective diagnosis algorithm as ISCD ${\beta}$-version. This system may help an oriental medical doctors to make a decision of Sasang constitutional diagnosis easily and correctly.

Structure and Expression of OsUBP6, an Ubiquitin-Specific Protease 6 Homolog in Rice (Oryza sativa L.)

  • Moon, Yea Kyung;Hong, Jong-Pil;Cho, Young-Chan;Yang, Sae-Jun;An, Gynheung;Kim, Woo Taek
    • Molecules and Cells
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    • v.28 no.5
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    • pp.463-472
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    • 2009
  • Although the possible cellular roles of several ubiquitin-specific proteases (UBPs) were identified in Arabidopsis, almost nothing is known about UBP homologs in rice, a monocot model plant. In this report, we searched the rice genome database (http://signal.salk.edu/cgi-bin/RiceGE) and identified 21 putative UBP family members (OsUBPs) in the rice genome. These OsUBP genes each contain a ubiquitin carboxyl-terminal hydrolase (UCH) domain with highly conserved Cys and His boxes and were subdivided into 9 groups based on their sequence identities and domain structures. RT-PCR analysis indicated that rice OsUBP genes are expressed at varying degrees in different rice tissues. We isolated a full-length cDNA clone for OsUBP6, which possesses not only a UCH domain, but also an N-terminal ubiquitin motif. Bacterially expressed OsUBP6 was capable of dismantling K48-linked tetra-ubiquitin chains in vitro. Quantitative real-time RT-PCR indicated that OsUBP6 is constitutively expressed in different tissues of rice plants. An in vivo targeting experiment showed that OsUBP6 is predominantly localized to the nucleus in onion epidermal cells. We also examined how knock-out of OsUBP6 affects developmental growth of rice plants. Although homozygous T3 osubp6 T-DNA insertion mutant seedlings displayed slower growth relative to wild type seedlings, mature mutant plants appeared to be normal. These results raise the possibility that loss of OsUBP6 is functionally compensated for by an as-yet unknown OsUBP homolog during later stages of development in rice plants.

Construction of Web-Based Database for Anisakis Research (고래회충 연구를 위한 웹기반 데이터베이스 구축)

  • Lee, Yong-Seok;Baek, Moon-Ki;Jo, Yong-Hun;Kang, Se-Won;Lee, Jae-Bong;Han, Yeon-Soo;Cha, Hee-Jae;Yu, Hak-Sun;Ock, Mee-Sun
    • Journal of Life Science
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    • v.20 no.3
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    • pp.411-415
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    • 2010
  • Anisakis simplex is one of the parasitic nematodes, and has a complex life cycle in crustaceans, fish, squid or whale. When people eat under-processed or raw fish, it causes anisakidosis and also plays a critical role in inducing serious allergic reactions in humans. However, no web-based database on A. simplex at the level of DNA or protein has been so far reported. In this context, we constructed a web-based database for Anisakis research. To build up the web-based database for Anisakis research, we proceeded with the following measures: First, sequences of order Ascaridida were downloaded and translated into the multifasta format which was stored as database for stand-alone BLAST. Second, all of the nucleotide and EST sequences were clustered and assembled. And EST sequences were translated into amino acid sequences for Nuclear Localization Signal prediction. In addition, we added the vector, E. coli, and repeat sequences into the database to confirm a potential contamination. The web-based database gave us several advantages. Only data that agrees with the nucleotide sequences directly related with the order Ascaridida can be found and retrieved when searching BLAST. It is also very convenient to confirm contamination when making the cDNA or genomic library from Anisakis. Furthermore, BLAST results on the Anisakis sequence information can be quickly accessed. Taken together, the Web-based database on A. simplex will be valuable in developing species specific PCR markers and in studying SNP in A. simplex-related researches in the future.

One-step Multiplex RT-PCR Method for Simultaneous Detection of Seed Transmissible Bacterium and Virus Occurring on Brassicaceae Crop Seeds (십자화과 작물 종자에서 종자전염 세균 및 바이러스 동시 검출을 위한 One-step Multiplex RT-PCR 방법)

  • Jeong, Kyu-Sik;Soh, Eun-Hee
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.52-58
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    • 2011
  • The aim of this research was to develop specific and sensitive PCR-based procedures for simultaneous detection of economically important plant pathogenic bacteria and seed borne virus in commercial Brassicaceae crop seeds, Xanthomonns campestris pv. campestris (Xcc) and Lettuce Mosaic Virus (LMV). Bacterial and virus diseases of Brassicaceae leaves are responsible for heavy losses. PCR with arbitral primers: selection of specific primers, performance of PCR with specific primers and determination of the threshold level for pathogens detection. To detect simultaneously the Xcc and LMV in commercial Brassicaceae crop seeds (lettuce, kohlrabi, radish, chinese cabbage and cabbage), two pairs of specific primer (LMV-F/R, Xcc-F/R) were synthesized by using primer-blast program (http://www.ncbi.nlm.nih.gov/tools/primer-blast/). The multiplex PCR for the two pathogens in Brassicaceae crop seeds could detect specifically without interference among primers and/or cDNA of other plant pathogens. The pathogen detection limit was determined at 1 ng of RNA extracted from pathogens. In the total PCR results for pathogen detection using commercial kohlrabi (10 varieties), lettuce (50 varieties), radish (20 varieties), chinese cabbage (20 varieties) and cabbage (20 varieties), LMV and Xcc were detected from 39 and 2 varieties, respectively. In the PCR result of lettuce, LMV and Xcc were simultaneously detected in 8 varieties.

Construction of web-based Database for Haliotis SNP (웹기반 전복류 (Haliotis) SNP 데이터베이스 구축)

  • Jeong, Ji-Eun;Lee, Jae-Bong;Kang, Se-Won;Baek, Moon-Ki;Han, Yeon-Soo;Choi, Tae-Jin;Kang, Jung-Ha;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.26 no.2
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    • pp.185-188
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    • 2010
  • The Web-based the genus Haliotis SNP database was constructed on the basis of Intel Server Platform ZSS130 dual Xeon 3.2 GHz cpu and Linux-based (Cent OS) operating system. Haliotis related sequences (2,830 nucleotide sequences, 9,102 EST sequences) were downloaded through NCBI taxonomy browser. In order to eliminate vector sequences, we conducted vector masking step using cross match software with vector sequence database. In addition, poly-A tails were removed using Trimmest software from EMBOSS package. The processed sequences were clustered and assembled by TGICL package (TIGR tools) equipped with CAP3 software. A web-based interface (Haliotis SNP Database, http://www.haliotis.or.kr) was developed to enable optimal use of the clustered assemblies. The Clustering Res. menu shows the contig sequences from the clustering, the alignment results and sequences from each cluster. And also we can compare any sequences with Haliotis related sequences in BLAST menu. The search menu is equipped with its own search engine so that it is possible to search all of the information in the database using the name of a gene, accession number and/or species name. Taken together, the Web-based SNP database for Haliotis will be valuable to develop SNPs of Haliotis in the future.