• The Plant Pathology Journal
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• 제35권1호
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• pp.51-62
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• 2019

A great variable response was observed when PP-3 and PP-J encumbered with 116 populations of root knot nematode (RKN) at two different temperatures ($25{\pm}2^{\circ}C$ and $30{\pm}2^{\circ}C$) and concentrations ($10^4$ and $10^5$ spores/ml). The PCR reaction amplified intergenic region between cytochrome oxidase subunit II gene (COII) and large subunit of rRNA gene (lrRNA) of the mitochondrial genome of different RKN species. The primer C2F3 and 1108 identified M. incognita with the highest frequency (52.6%) followed by M. javanica (36.8%) and M. arenaria (10.5%). The sizes of PCR products were 1.7 kb for M. incognita and M. javanica populations while populations of M. arenaria produced 1.1 kb fragment. The digestion with Hinf I yielded three different fragment length patterns on 1.5 % agarose gel. From current research it is concluded that intra-Meloidogyne genetic variability exist in RKN populations which have better encumbrance with P. penetrans.

• 대한본초학회지
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• 제27권6호
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• pp.15-21
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• 2012

Objective : Lonicera japonica THUNB. a traditional herbal medicine, has been commonly used anti-inflammatory disease. It has been very complicated with respect to its sources on the market. The significant selection of medicine depends on its origin. However, it is difficult to discrimination criteria for confirming L. japonica authenticity using the senses. This study was performed to determine the discriminant analysis of L. japonica and L. confusa. Methods : The identification of L. japonica and L. confusa were performed by the classification and identification committee of the national center for standardization of herbal medicines. And we examined its differences using HPLC and genetic marker analysis. Results : The analytical pattern of High Performance Liquid Chromatography was determined from the corresponding peak curves ((E)-aldosecologanin, chlorogenic acid, luteolin 7-O-glucoside, sweroside). For L. japonica, additional unknown peaks were detected at 13.8 min, 20.6 min, and 36.9 min. And, we developed genetic marker using the the tRNA-Leu gene, trnL-trnF intergenic spacer and tRNA-Phe region of chloroplast DNA. By the method, 164 bp PCR product amplified from L. confusa was distinguished into L. japonica and L. confusa efficiently. Conclusion : Base on these results, two techniques provide effective approaches to distinguish L. japonica from L. confusa.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2021년도 춘계학술대회
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• pp.16-16
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• 2021

To distinguish manchurian clematis, Clematis terniflora var. mandshurica and three-leaf clematis, C. apiifolia, we collected 9 nuclear ITS sequences and two sequences of trnQ-trnH intergenic spacer and trnH region in GenBank database. Those sequences were aligned to find differences between those of C. terniflora var. mandshurica and C. apiifolia. Two primer pairs were newly designed base on the differences between two species and conducted multiplex PCR. The size of amplified fragments using generated primers were 380 base pairs (only three-leaf clematis) and 189 base pairs (both species). This genetic marker based on PCR is useful to discrimination of C. terniflora var. mandshurica and C. apiifolia

• BMB Reports
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• 제40권5호
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• pp.740-748
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• 2007

To gain insight into the structure and function of repressor proteins of bacteriophages of gram-positive bacteria, repressor of temperate Staphylococcus aureus phage ${\phi}11$ was undertaken as a model system here and purified as an N-terminal histidine-tagged variant (His-CI) by affinity chromatography. A ~19 kDa protein copurified with intact His-CI (~ 30 kDa) at low level was resulted most possibly due to partial cleavage at its Ala-Gly site. At ~10 nM and higher concentrations, His-CI forms significant amount of dimers in solution. There are two repressor binding sites in ${\phi}11$ cI-cro intergenic region and binding to two sites occurs possibly by a cooperative manner. Two sites dissected by HincII digestion were designated operators $O_L$ and $O_R$, respectively. Equilibrium binding studies indicate that His-CI binds to $O_R$ with a little more strongly than $O_L$ and binding species is probably dimeric in nature. Interestingly His-CI binding affinity reduces drastically at elevated temperatures ($32-42^{\circ}C$). Both $O_L$ and $O_R$ harbor a nearly identical inverted repeat and studies show that ${\phi}11$ repressor binds to each repeat efficiently. Additional analyses indicate that ${\phi}11$ repressor, like $\lambda$ repressor, harbors an N-terminal domain and a C-terminal domain which are separated by a hinge region. Secondary structure of ${\phi}11$ CI even nearly resembles to that of $\lambda$ phage repressor though they differ at sequence level. The putative N-terminal HTH (helix-turn-helix) motif of ${\phi}11$ repressor belongs to the HTH -XRE-family of proteins and shows significant identity to the HTH motifs of some proteins of evolutionary distant organisms but not to HTH motifs of most S. aureus phage repressors.

• Journal of Plant Biotechnology
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• 제44권2호
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• pp.135-141
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• 2017

구지뽕 나무(Cudrania tricuspidata Bureau)는 일반적으로 이용되는 대표적인 다년생의 약용식물이다. 최근 국제적 추세에 따라 자국의 유전자원의 발굴, 보존 등이 강화됨에 따라 인접국가와 국내 자생 꾸지뽕 계통을 판별 할 수 있는 기준설정에 관한 연구의 필요성이 대두되고 있지만, 분자생물학적 판별 기술의 개발은 아직 미흡한 실정이다. 본 연구에서는 국내 토종과 중국 계통의 구지뽕 나무의 기원을 판별하기 위해 엽록체에 존재하는 trnL-trnF 유전자단편에서 SNP를 이용한 판별 프라이머를 확보하였으며 이를 보완하여 보다 신속하게 판별하기 위하여 ARMS-PCR기술을 이용한 판별마커와 그 조건을 확립하였다. 그러므로, 본 연구에서 개발된 SNP 마커는 다양한 지역에서 서식하는 구지뽕 계통들의 신속한 확인을 위해 매우 유용하게 이용될 것으로 생각된다.

• Journal of Plant Biotechnology
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• 제45권2호
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• pp.102-109
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• 2018

엉겅퀴는 일반적으로 이용되는 대표적인 다년생의 약용식물이다. 최근 국제적 추세에 따라 자국의 유전자원의 발굴, 보존 등이 강화됨에 따라 인접국가와 국내 자생 엉겅퀴 계통을 판별 할 수 있는 기준 설정에 관한 연구의 필요성이 대두되고 있지만, 분자생물학적 판별 기술의 개발은 아직 미흡한 실정이다. 본 연구에서는 국내 토종과 해외 유래 엉겅퀴종의 기원을 판별하기 위해 핵의 리보솜에 존재하는 ITS 유전자단편에서 SNP를 이용한 판별 프라이머를 확보하였으며, 이를 보완하여 보다 신속하게 판별하기 위하여 ARMS-PCR 및 HRM 기술을 이용한 판별 마커와 그 조건을 확립하였다. 또한, 국내 종 특이적 프라이머들을 이용한 정량적 PCR 분석방법을 이용해 두 가지 종의 genomic DNA의 혼합 여부를 판별하였다. 그러므로, 본 연구에서 개발된 SNP 마커는 다양한 지역 또는 국가에서 서식하는 엉겅퀴 종들의 신속한 확인을 위해 매우 유용하게 이용될 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1841-1848
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• 2006

Insights into gene expression have the potential for improvement of antibiotic yield and the development of robust production hosts for use in recombinant biomolecule production. $Cubicin^{TM}$ (daptomycin for injection) is a recently approved antibiotic active against many Gram(+) pathogens, including those resistant to methicillin, vancomycin, and fluoroquinolones. Daptomycin is produced as a secondary metabolite by Streptomyces roseosporus. A 128 kb region of DNA including the daptomycin biosynthetic gene cluster (dpt) has been cloned. and sequenced. Using a selected array of nucleic acid probes representing this region, we compared the expression levels of the dpt genes between S. roseosporus wild-type (WT) and derived S. roseosporus high-producer of daptomycin (HP). We observed that the majority of the biosynthetic genes were upregulated in HP compared with WT; a total of 12 genes, including those encoding daptomycin synthetase, showed consistently and significantly higher expression levels, at least 5-fold, in HP compared with WT. In contrast, some genes, flanking the dpt cluster, were expressed at higher levels in the WT strain. The expression of housekeeping genes such as S. roseosporus rpsL, rpsG, and 16S (positive controls) and presumptive intergenic regions in the dpt cluster (negative control) were identical in the two strains. In addition, we compared transcription during the early, mid-log, and early-stationary phases of growth in the HP strain. The same set of genes was upregulated and downregulated under all conditions examined; housekeeping genes showed no relative change in expression level over the periods of growth tested. Analyses of this type would be of value in studies of strain improvement and also for the identification of gene regulation processes that are important for secondary metabolite production.

• Molecules and Cells
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• 제27권3호
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• pp.365-381
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• 2009

The chloroplast DNA sequences of Megaleranthis saniculifolia, an endemic and monotypic endangered plant species, were completed in this study (GenBank FJ597983). The genome is 159,924 bp in length. It harbors a pair of IR regions consisting of 26,608 bp each. The lengths of the LSC and SSC regions are 88,326 bp and 18,382 bp, respectively. The structural organizations, gene and intron contents, gene orders, AT contents, codon usages, and transcription units of the Megaleranthis chloroplast genome are similar to those of typical land plant cp DNAs. However, the detailed features of Megaleranthis chloroplast genomes are substantially different from that of Ranunculus, which belongs to the same family, the Ranunculaceae. First, the Megaleranthis cp DNA was 4,797 bp longer than that of Ranunculus due to an expanded IR region into the SSC region and duplicated sequence elements in several spacer regions of the Megaleranthis cp genome. Second, the chloroplast genomes of Megaleranthis and Ranunculus evidence 5.6% sequence divergence in the coding regions, 8.9% sequence divergence in the intron regions, and 18.7% sequence divergence in the intergenic spacer regions, respectively. In both the coding and noncoding regions, average nucleotide substitution rates differed markedly, depending on the genome position. Our data strongly implicate the positional effects of the evolutionary modes of chloroplast genes. The genes evidencing higher levels of base substitutions also have higher incidences of indel mutations and low Ka/Ks ratios. A total of 54 simple sequence repeat loci were identified from the Megaleranthis cp genome. The existence of rich cp SSR loci in the Megaleranthis cp genome provides a rare opportunity to study the population genetic structures of this endangered species. Our phylogenetic trees based on the two independent markers, the nuclear ITS and chloroplast MatK sequences, strongly support the inclusion of the Megaleranthis to the Trollius. Therefore, our molecular trees support Ohwi's original treatment of Megaleranthis saniculifolia to Trollius chosenensis Ohwi.

• Journal of Plant Biotechnology
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• 제43권2호
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• pp.164-173
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• 2016

수박의 형태적 변이의 유전적 원인을 분석해 보기 위해 8개 계통에서 re-sequencing을 수행하였다. 유전적 변이의 수는 염색체에 따라 다르게 나왔으며 발견된 SNP의 약 12.9%만이 유전자내에서 발견되었고 나머지는 프로모터나 유전자 사이의 지역에서 발견되었다. SNP 밀도에 대한 분석 결과 염색체 6번의 말단지역에 변이가 집중되어 있는 것을 알 수 있었다. 또한 염색체 10과 11번에 잘 보존된 지역을 발견하였다. Pathway 분석을 통해 DIMBOA(일종의 항생제)-glucoside 분해 대사가 계통간 가장 차이나는 것으로 확인되었으며 이는 각 계통의 병저항성에서 차이가날 가능성을 시사하는 것이다. 당대사 관련 유전자 변이를 분석한 결과 alpha-galactosidase 유전자에 가장 변이가 많은 것으로 밝혀졌다. 이러한 연구 결과는 육종을 분자수준에서 이해하는 데 도움을 줄 것으로 생각한다.

• Horticulture, Environment, and Biotechnology : HEB
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• 제59권6호
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• pp.865-873
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• 2018

Allium ochotense and Allium microdictyon are commonly known as 'Mountain garlic' and are popular, economically important species in many countries such as Korea, China, and Mongolia. Their leaves are used as culinary side dishes and in traditional medicines. In Korea, these two species are at risk of extinction due to damage to their natural habitat and thus, conservation and breeding programs are needed. However, their identification relies mostly on morphological data, which is limited and until recently, led to classifying these two species under A. victorialis. In the present study, a simple and reliable method of molecular identification was developed to distinguish A. ochotense from A. microdictyon that targets four barcoding regions: the internal transcribed spacer (ITS), the maturase K gene (matK), the chloroplast psbA-trnH intergenic region, and the ribulose-bisphosphate carboxylase large subunit gene (rbcL). Single nucleotide polymorphisms (SNPs) were found in ITS and matK regions, and species-specific primers were designed based solely on the SNP at position 680 of the ITS region that could differentiate A. ochotense from A. microdictyon. Using these primers in amplification refractory mutation system (ARMS)-PCR, A. ochotense, and A. microdictyon could be simultaneously and efficiently distinguished. This study is the first to report a simple, rapid, and efficient method for discriminating A. ochotense and A. microdictyon, indicating the utility of species-specific markers in the development of conservation and breeding programs.