• 한국자원식물학회지
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• 제17권2호
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• pp.116-121
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• 2004

고려 인삼중 폐포와 4등급 이하를 유발시키는 90％이상이 적변삼이라고 불리는 인삼의 표피 색택이 붉은 삼이 그 원인이다. 이러한 적변삼은 미국삼보다는 고려 인삼에 서 다량 발견되는 바, 적변은 유전적 요인이 있다고 사료된다. 그러므로 이 연구의 목적은 RT-PCR을 이용하여 인삼적병에 내성을 가지는 유전자를 탐색하기 위하여 실시되었다. 고려인삼 3년근 1개체 중에서 적변이 발생된 부위와 건전 부위의 RNA를 추출하여 형성된 cDNA를 여러개의 random primer를 사용하여 PCR 증폭을 한 결과 정상 부위의 cDNA에서 발견되지 않는 band가 적변삼의 부위에서 발견되었다. 따라서 band가 형성된 부위의 유전자가 적변과 관련될 가능성이 있는 것으로 사료되고 이러한 유전자는 향후 염기서열을 분석하여 어떠한 유전자인지 판명을 하여야 하며 적변관련 유전자이면 선발마커로서 사용되고 또한 형질전환을 통한 적변내성 인삼계통을 육성할 수 있으며, 만약 적변과 관련이 없는 유전자로 판명된다면 더 많은 primer를 사용하여 적변관련 유전자를 탐색해야 할 것이다.

• The Plant Pathology Journal
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• 제27권4호
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• pp.367-371
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• 2011

Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.

• The Plant Pathology Journal
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• 제26권4호
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• pp.409-416
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• 2010

Fusarium species, a large group of plant pathogens, potentially pose quarantine concerns worldwide. Here, we focus on the development of a method for detecting four Fusarium species in quarantined plants in Korea: F. solani f. sp. cucurbitae, F. stilboides, F. redolens, and F. semitectum var. majus. Species-specific primers were designed from the nucleotide sequences of either the translation elongation factor-1 alpha (TEF1) gene or RNA polymerase II subunit (RPB2) gene. Two different primer sets derived from TEF1, all specific to F. solani f. sp. cucurbitae, were able to differentiate the two races (1 and 2) of this species. A set of nested primers for each race was designed to confirm the PCR results. Similarly, two primer sets derived from RPB2 successfully amplified specific fragments from five F. stilboides isolates grouped within a single phylogenetic clade. A specific TEF1 primer set amplified a DNA fragment from only four of the 12 F. redolens strains examined, which were grouped within a single phylogenetic clade. All of the F. semitectum var. majus isolates could be specifically detected with a single RPB2 primer set. The specificity of the primer sets developed here was confirmed using a total of 130 Fusarium isolates.

• Journal of Microbiology and Biotechnology
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• 제17권10호
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• pp.1708-1711
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• 2007

The Streptomyces coelicolor M145 genome harbors six copies of divergent rRNA operons that differ at ${\sim}0.2%$ and ${\sim}0.6%$ of the nucleotide positions in small subunit (SSU) and large subunit (LSD) rRNA genes, respectively. When these rRNA genes are expressed, a single cell may harbor three different kinds of SSU rRNA and five kinds of LSU rRNA. Primer extension analyses revealed that all of the heterogeneous rRNA molecules are expressed and assembled into ribosomes. This finding and the maintenance of the intragenomic variability of rRNA operons imply the existence of functional divergence of rRNA species in this developmentally complex microorgamsm.

• The Plant Pathology Journal
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• 제21권4호
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• pp.343-348
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• 2005

The partial nucleotide sequences of the genomic dsRNA mycoviruses infecting Pleurotus ostreatus (isolates ASI2596, ASI2597, and Bupyungbokhoe) and Agaricus blazei Murrill were determined and compared with those of the other dsRNA mycoviruses. Partial nucleotide sequences of the purified dsRNA from ASI2596 and ASI2597 revealed RNA-dependent RNA polymerase sequences that are closely related to Oyster mushroom isometric virus 2, while nucleotide sequences and the deduced amino acid sequence from dsRNA mycovirus infecting Agaricus blazei did not show any significant homology to the other dsRNA mycoviruses. Specific primers were designed for RT-PCR detection of these dsRNA viruses and were found to specifically detect each dsRNA virus. Northern blot analysis confirmed the homogeneity of RT-PCR products to each purified dsRNA. Altogether, our results suggest that these virus-specific primer sets can be employed for the specific detection of each dsRNA mycovirus in infected mushrooms.

• Parasites, Hosts and Diseases
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• 제35권3호
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• pp.203-210
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• 1997

이질아메바 병원성 분리주에서 특이적으로 발현되는 mRNA를 동정하고자 differential display reverse transcription-polymerase chain reaction(DDRT-PCR)을 수행하여 병원성 특이 증폭산물을 확인하였다. 한국인에서 검출한 이질아메바 병원성 분리주 YS-27과 Entamoeba dispar분리주인 S 16으로부터 정제한 mRAN를 주형으로 11개의 arbitrary primer와 3개의 one base anchored $oligo-dT_{11}M$(M: A, C 또는 G)의 조합을 이용, DDRT-PCR을 실시한 결과 31개의 분획이 YS-27주에서만 증폭된 것으로 확인되었다. 이 331개 DNA 중 21개는 cysteine proteinase 유전자와 상동성을 나타내었다. YS-27주로부터 제작된 cDNA library를 나머지 DNA를 탐침으로 사용, 검색하여 최종 4개의 clone을 얻었다. 이 4개의 clone을 이용, immunoscreening을 수행한 결과, 이 clone들은 이질아메바 감염자 혈청과 양성반응을 나타내고 있었다.

• Animal cells and systems
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• 제1권2호
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• pp.363-370
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• 1997

RNase MRP is a ribonucleoprotein complex with a site-specific endonuclease activity. Its original substrate for cleavage is the small mitochondrial RNA near the mitochondrial DNA replication origin, thus it was proposed to generate the primer for mtDNA replication. Recently, it has been shown to have another substrate in the nucleus, such as pre-S.8S ribosomal RNA in nucleolus. The gene for the RNA component of RNase MRP (MRP RNA) was found to be encoded by the nucleus genome, suggesting an interesting intracellular trafficking of MRP RNA to both mitochondria and nucleolus after transcription in nucleus. In this study, genomic DNA encoding MRP RNA was microinjected into the nucleus of Xenopus oocytes, to analyze promoter regions involved in the transcription. It showed that the proximal sequence element and TATA box are important for basal level transcription; octamer motif and Sp1 binding sites are for elevated level transcription. Most of Xenopus MRP RNA was exported out to the cytoplasm following transcription in the nucleus. Utilizing various hybrid constructs, export of MRP RNA was found to be regulated by the promoter and the 5' half of the coding region of the gene. Interestingly, the transcription in nucleus seems to be coupled to the export of MRP RNA to cytoplasm. Intracellular transport of injected MRP RNA can be easily visualized by whole-mount in situ hybridization following microinjection; it also shows possible intra-nuclear sites for transcription and export of MRP RNA.

• BMB Reports
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• 제49권11호
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• pp.612-616
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• 2016

CD44 pre-mRNA includes 20 exons, of which exons 1-5 ($C_1-C_5$) and exons 16-20 ($C_6-C_{10}$) are constant exons, whereas exons 6-15 ($V_1-V_{10}$) are variant exons. $V_6$-exon-containing isoforms have been known to be implicated in tumor cell invasion and metastasis. In the present study, we performed a SR protein screen for CD44 $V_6$ splicing using overexpression and lentivirus-mediated shRNA treatment. Using a CD44 $V_6$ minigene, we demonstrate that increased SRSF3 and SRSF4 expression do not affect $V_6$ splicing, but increased expression of SRSF1, SRSF6 and SRSF9 significantly inhibit $V_6$ splicing. In addition, using a constitutive exon-specific primer set, we could not detect alterations of CD44 splicing after SR protein-targeting shRNA treatment. However, using a $V_6$ specific primer, we identified that reduced SRSF2 expression significantly reduced the $V_6$ isoform, but increased $V_{6-10}$ and $V_{6,8-10}$ isoforms. Our results indicate that SR proteins are important regulatory proteins for CD44 $V_6$ splicing.

• 생명과학회지
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• 제12권2호
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• pp.158-163
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• 2002

남조류 Microcystis aeruginosa를 무균적으로 분리하기 위해 낙동강 물금지역의 수화를 멸균 증류수로 vortex 전처리를 하였으며, 세균제거 및 무균상태를 계속 유지하기 위하여 항생물질(ampicillin 150 $\mu$g/$m\ell$, neomycin 25 $\mu$g/$m\ell$)을 배지에 첨가하고, 독립 집락으로 형성시켜 오염 기회를 줄이기 위하여 0.7% agarose로 고형화시킨 CB고체배지에서 3$0^{\circ}C$, 40 $\mu$mol m$^{-2}$ s$^{-1}$ 광 조건으로 배양하였다. 그 결과 분리되어진 26개의 Microcystis aeruginosa colony 중 3개의 무균 균주만이 확보되었다. 3개의 무균균주를 16S rRNA primer를 이용하여 PCR 증폭한 결과 M. aeruginosa AF 139292와 99.5에서 100%의 상동성을 가지는 것으로 나타났다.

• 한국동물위생학회지
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• 제35권4호
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• pp.289-294
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• 2012

Mycoplasmas are highly fastidious bacteria, difficult to culture and slow growing. Infections with Mycoplasma species can cause a variety of problems in living organisms and in vitro cell cultures. In this study, we investigated the usefulness of a genus-specific consensus PCR analysis method to detect Mycoplasma species. The developed consensus primer pairs MycoF and MycoR were designed specifically to amplify the 16S ribosomal RNA gene (rRNA) of Mycoplasma species by the optimized PCR system. The developed consensus PCR system effectively amplified 215 bp of Mycoplasma genus-specific region of 16S rRNA. In conclusion, we recommend this consensus PCR for monitoring Mycoplasma species in animals, human and cell culture system.