• 대한바이러스학회지
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• 제27권1호
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• pp.1-8
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• 1997

Human caliciviruses (HuCVs) cause sporadic cases and outbreaks of acute gastroenteritis (AGE). Three major genogroups of HuCVs have been described including the Norwalk virus (NV)-, the Snow Mountain virus (SMA)-, and the Sapporo-genogroups. This study describes the detection and genetic variation of HuCVs from hospitalized infants with AGE in Korea by RT-PCR and sequencing. The cDNA fragments of 206 to 470bp corresponding to the region of 3 primer pairs (36/35, 35/51 or 3/51) in the polymerase region of NV were generated. Of 185 stools screened, 8% were positive by RT-PCR and their sequences showed that all strains contained the GLPSG and YGDD motifs which are conserved for HuCVs. Amino acid (aa) sequence analysis showed that these strains can be divided into 3 major genogroups. High conservation was observed in that one strain shares 100% of aa sequence with Southampton virus, another shares 99% with the Sapporo virus, and six strains share 90 to 95% with Snow Mountain virus. However, significant sequence variation was also found in other strains. This study indicates that all major genogroups of HuCVs are circulating in Korea.

• Journal of Microbiology
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• 제34권1호
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• pp.15-22
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• 1996

The cloned X-bacterial gene (groEx) which is analogous to groE of E. Coli strongly expressed in E. coli when grown at the temperature 27.deg. C or higher without having to add any inducers. By S1-nuclease mapping, primer extension analysis and site directed mutagenesis, we found 4 promoters in the gene. Among them two promoters located at 5'-extended region of the gene are homologous to the promoters found in groE family of heat-shock genes ; they are , .sigma.$^{32}$ factor-dependent P1 promotor and .delta$^{70}$factor-dependet P2 promoter. The other two promoters found within the coding region of groESx were P3, 5'-TTGGCG-(18 bases)-AATACT-3' and P4, 5'-TTGGCA-(19 bases)-TAAGT which overlapped within 49 bases. These unique intragenic .delta.$^{70}$-dependent promoters are the first to be cloned and characterized in groE analogous heat-shock genes so far. These P3 and P4 promoters appeared to be responsible for the strong expression of GroElx in X-bacteria in vivo.

• Biomolecules & Therapeutics
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• 제16권3호
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• pp.184-189
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• 2008

The proto-oncogene protein DEK has been implicated in various human disease including cancer. We have shown that DEK induces caspase-dependent apoptosis in Drosophila by regulating histone acetylation. Reverse transcription-polymerase chain reaction (RT-PCR) method based on annealing control primers was used to screen and identify differentially expressed genes (DEGs) in DEK overexpressed HeLa cells. Among the genes identified, clusterin and fibrillarin have major role in apoptosis pathway regulation. TFIIIC and RPS24 are implicated in HAT mediated transcriptional initiation and cololectal cancer, respectively. To further analyze DEK's role in apoptosis, multiplex PCR was performed. Caspase-3, -7, and -10 and proapoptotic gene bid were newly identified as possible target genes regulated by DEK expression.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2002년도 추계학술대회초록집
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• pp.139-139
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• 2002

돼지 설사유발 칼리시 바이러스(Porcine enteric calicivirus: PECV)도 자돈에서 설사를 일으키는 바이러스로 이미 알려졌다. RT-PCR과 nested PCR을 이용하여 국내 양돈장에서 PECV의 발생을 조사하고자 본 연구를 시도하였다. 설사 분변은 경기, 충남, 전북, 전남과 제주지역에 분포한 31개의 농장 102마리의 자돈에서 채취하여 의뢰된 것을 조사하였다. RT-PCR 과 nested PCR 을 위하여 RNA dependent RNA Polymerase (RDRP) 부위와 capsid 부위에서 각각 2 쌍의 primer를 작성하였다. RDRP 부위에서 RT-PCR을 시행했던 바, 3마리 (2.9％) 에서, nested PCR에서는 18마리 (17.6％)에서 양성반응이 나왔으며 capsid 부위에서 RT-PCR 결과 5마리 (4.9％), nested PCR에서는 18마리(17.6％)가 양성반응으로 확인되었다. 본 연구를 통하여 PECV가 국내에서 돼지 설사를 일으키는 주요 원인체 중 하나라는 것이 밝혀졌으며, nested PCR 기법이 돼지 설사분변에서 PECV를 검출하는 좋은 진단방법이었다.

• 한국산림과학회지
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• 제96권2호
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• pp.125-130
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• 2007

The principal morphological characters used for identification of hydrangea cultivars are often dependent on agroclimatic conditions. Furthermore, information on the selection or the genetic background of the hydrangea breeding is so rare that a molecular marker system for cultivar identification is needed. Amplified fragment length polymorphism (AFLP) markers were employed for fingerprinting Hydrangea macrophylla cultivars and candidate cultivars of H. macrophylla selected in Korea. One AFLP primer combination was sufficient to distinguish 17 H. macrophylla cultivars and 4 candidate cultivars. The profile of 19 loci that can minimize the error of amplification peak detection was constructed. AFLP markers were efficient for identification, estimation of genetic distances between cultivars, and cultivar discrimination. Based on the observed AFLP markers, genetic relationship was reconstructed by the UPGMA method. Seventeen H. macrophylla cultivars and H. macrophylla for. normalis formed a major cluster, and candidate cultivars selected in Korea formed another cluster.

• The Plant Pathology Journal
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• 제20권3호
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• pp.200-205
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• 2004

The partial nucleotide sequences of the genomic dsRNA mycovirus infecting Pleurotus ostreatus isolates ASI2223 and Suhan were determined and compared with those of mycoviruses belonging to partitiviruses and totiviruses. Partial nucleotide sequences of the purified dsRNA from ASI2223 and Suhan showed RNA-dependent RNA polymerase sequences that are closely related to those of partitiviruses, including Fusarium poae virus 1, Fusarium solani virus, Rhizoctoniasolani virus, Discula destructiva virus 2, and Oyster mushroom isometric virus 2. Specific primers were designed for RT-PCR detection of dsRNA viruses from the P. ostreatus isolate ASI2223 and Suhan. Two virus specific primer sets were found to specifically detect each virus among six sets of designed oligonucleotide primers. Collectively, these results suggest that dsRNA mycoviruses from P. ostreatus isolates ASI2223 and Suhan belong to the family Partitiviridae, although, they are not the same virus species. Our results also suggest that these virus-specific primer sets can be employed for the specific detection of each viral sequence in infected tissues.

• 대한기계학회논문집A
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• 제31권7호
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• pp.808-815
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• 2007

The optical performance of the mirror fur satellite camera is highly dependent on the adhesive properties between the mirror and its support. Therefore, in order to design a mirror with high optical performance, the mechanical properties of adhesives should be well defined. In this research, the mechanical properties of three kinds of space adhesives are studied. In case of the materials which show nearly incompressible behavior such as space adhesives, it is important to measure shear modulus which governs deviatoric stress components. Also the experiment should be performed in circumstances similar to real manufacturing process of mirror, because extra factors such as size effects, the adhesion effects of primer and reactions between adhesive and primer affect the properties of adhesive regions. In this research shear moduli of the adhesives are determined by using a single lap adhesively bonded joint. For the shear tests, several temperatures have been selected from $-20^{\circ}C$ to $55^{\circ}C$ which is operating temperature range of the adhesive. In the case of linear behavior materials, shear moduli are calculated through a linear curve fitting. Shear stress-strain relation is obtained by using an exponential curve fitting for material which shows non-linear behavior. The shear modulus of each adhesive is expressed as a function of temperature. Characteristics and adaptability of the adhesives are discussed regarding their temperature sensitivity.

• Molecules and Cells
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• 제26권3호
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• pp.250-257
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• 2008

Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.

• 한국균학회지
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• 제20권4호
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• pp.315-323
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• 1992

재현성 있는 RAED marker들의 증폭을 위해서 PCR에 영향을 주는 조건들에 대해 조사를 하였다. 그 결과 약 15 ng의 DNA가 효과적인 PCR에 적합한 양이었으며 PCR에 사용되는 성분(reaction component)들의 농도가 PCR 결과에 있어서 상호의존관계에 있었고 DNA 용액에 포함되어 있는 RNA가 DNA 증폭을 방해하는 작용을 하였다. $25\;{\mu}l$의 PCR 반응용액에 30ng의 10-mer primer, $200\;{\mu}M$ dNTP, 0.001% gelatin, 1.5 mM $MgCl_2$, 10 mM Tris-Cl(pH 8.8), 50 mM KCl, 0.1%, Triton X-100, 2units의 Taq DNA polymerase, 그리고 RNA를 제거한 15 ng의 DNA를 사용한 결과 가장 재현성있는 RAPD marker들이 증폭되었다.

• Journal of Crop Science and Biotechnology
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• 제11권2호
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• pp.127-134
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• 2008

Tobamoviruses are the major viral pathogens of tomato and bell pepper. The preliminary results showed that Acibenzolar-Smethyl(ASM; S-methylbenzo(1,2,3) thiadiazole-7-carbothiate) pre-treatment to tomato and tobacco plants reduces the concentration of Tomato mosaic tobamovirus(ToMV) and Tobacco mosaic tobamovirus(TMV) in tomato and bell pepper seedlings, respectively. Pre-treatment of the indicator plant(Nicotiana glutinosa) with the ASM followed by challenge inoculation with tobamoviruses produced a reduced number and size of local lesions(67 and 79% protection over control to TMV and ToMV inoculation, respectively). In order to understand the mechanism of resistance the gene expression profiles of antiviral genes was examined. RT-PCR products showed higher expression of two viral resistance genes viz., alternative oxidase(AOX) and RNA dependent RNA polymerase(RdRp) in the upper leaves of the ASM-treated tomato plants challenge inoculation with ToMV. Further, the viral concentration was also quantified in the upper leaves by reverse transcription PCR using specific primer for movement protein of ToMV, as well as ELISA by using antisera against tobamoviruses. The results provided additional evidence that ASM pre-treatment reduced the viral movement to upper leaves. The results suggest that expressions of viral resistance genes in the host are the key component in the resistance against ToMV in the inducer-treated tomato plants.