• 대한한의학회지
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• 제27권2호
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• pp.225-231
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• 2006

Objectives : This study was performed to determine if unknown species of antler samples could be identified by genetic distance methods. Methods : The DNAs of 4 antler samples were extracted, amplified by PCR, and sequenced. The DNAs of antlers were identified by genetic distance. Genetic distance method was made using MEGA software (Molecular Evolutionary Genetics Analysis, 3.1). Results : By genetic distance methods, all 4 antler samples were closest to Cervus elaphus nelsoni among Cervus species. Conclusion : These results suggest that genetic distance methods might be used as a tool for the identification of Cervus species.

• International Journal of Oral Biology
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• 제35권1호
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• pp.21-25
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• 2010

This study was undertaken to develop species-specific forward and universal reverse PCR primers for the detection of Streptococcus sobrinus. These primers target the variable regions of the 16S ribosomal RNA coding gene (rDNA) and their specificity was tested against 10 strains of S. sobrinus strains and 20 different species of oral bacteria using serial dilutions of the purified genomic DNA of S. sobrinus ATCC $33478^T$. Our data show that species-specific amplicons were obtained from all the S. sobrinus strains tested but not from other species. Both direct and nested PCR could detect as little as 400 pg and 4 fg of genomic DNA from S. sobrinus ATCC $33478^T$, respectively. This result suggests that these PCR primers are highly specific and sensitive and applicable to the detection of S. sobrinus.

• 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.

• 한국양식학회지
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• 제21권3호
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• pp.139-145
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• 2008

성게의 형태학적 분류는 그것의 형질적 변이에 의하여 어려움이 많다. 본 연구에서는 말똥성게, 둥근성게, 보라성게, 분홍성게와 동해안에서 포획된 미확인 성게 4종의 형태형질 비교와 계통유연관계를 조사하였다. 성게의 생식소로부터 genomic DNA를 분리한 후, PCR 방법을 통하여 mitochondrial 12S rDNA 유전자의 염기서열을 분석하였다. 둥근성게과의 말똥성게, 둥근성게, 만두성게과의 보라성게, 주발성게과의 분홍성게의 mitochondrial 12S rDNA의 염기서열은 미확인 성게종들의 그것과 85.9-93.9%의 상동성을 나타내었다. 한편, 미확인 성게종들은 새치성게의 mitochondrial 12S rDNA의 일부 염기서열과 99.8%의 높은 상동성을 보였으며, 각 개체의 mitochondrial 12S rDNA를 통한 분자계통수 분석에 의해서 미확인 성게들은 새치성게의 형태적 변이로 판단된다.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.487-488
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• 2000

Genomic DNA was isolated from the marine microalgae representing genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA as arbitrary primers. The electrophoretc analysis of PCR-RAPD products showed hig levels of variation between different genus and little variation between different species. Outer of these primers, 6 generated 248 highly reproducible RAPD markers, producing almost seven polymorphic bands per primers. The degree of similarity frequency between Chaetoceros gracilis and Chaetoceros calcitrans species showed 90％ as calculated by sharing analysis. The RAPD polymorphism generated by this primer may be used as a genetic marker for genus or species identification in important marine microalgae. (omitted)

• BMB Reports
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• 제37권2호
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• pp.144-147
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• 2004

Molecular amplification and sequencing of genomic DNA that encodes camel polyubiquitin (PUBC1) was performed by a polymerase chain reaction (PCR) using various sets of primers. The amplification generated a number of DNA fragments, which were sequenced and compared with the polyubiquitin coding sequences of various species. One DNA fragment that conformed to 325 bp was found to be 95 and 88% homologous to the sequences of human polyubiquitin B and C, respectively. The DNA translated into 108 amino acids that corresponded to two fused units of ubiquitin with no intervening sequence, which indicates that it is a polyubiquitin and contains at least two units of ubiquitin. Although, variations were found in the nucleotide sequence when compared to those of other species, the amino acid sequence was 100% homologous to the polyubiquitin sequences of humans, mice, and rats. This is the first report of the polyubiquitin DNA coding sequence and its corresponding amino acid sequence from camels, amplified using direct genomic DNA preparations.

• 원예과학기술지
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• 제35권2호
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• pp.149-164
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• 2017

Next generation sequencing (NGS) technologies have led to the rapid accumulation of genome sequences through whole-genome sequencing and re-sequencing of crop species. Genomic resources provide the opportunity for a new revolution in plant breeding by facilitating the dissection of complex traits. Among vegetable crops, reference genomes have been sequenced and assembled for several species in the Solanaceae and Cucurbitaceae families, including tomato, pepper, cucumber, watermelon, and melon. These reference genomes have been leveraged for re-sequencing of diverse germplasm collections to explore genome-wide sequence variations, especially single nucleotide polymorphisms (SNPs). The use of genome-wide SNPs and high-throughput genotyping methods has led to the development of new strategies for dissecting complex quantitative traits, such as genome-wide association study (GWAS). In addition, the use of multi-parent populations, including nested association mapping (NAM) and multiparent advanced generation intercross (MAGIC) populations, has helped increase the accuracy of quantitative trait loci (QTL) detection. Consequently, a number of QTL have been discovered for agronomically important traits, such as disease resistance and fruit traits, with high mapping resolution. The molecular markers for these QTL represent a useful resource for enhancing selection efficiency via marker-assisted selection (MAS) in vegetable breeding programs. In this review, we discuss current genomic resources and marker-trait association analysis to facilitate genome-assisted breeding in vegetable species in the Solanaceae and Cucurbitaceae families.

• Mycobiology
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• 제42권4호
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• pp.311-316
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• 2014

Lichen studies, including biodiversity, phylogenetic relationships, and conservation concerns require definitive species identification, however many lichens can be challenging to identify at the species level. Molecular techniques have shown efficacy in discriminating among lichen taxa, however, obtaining genomic DNA from herbarium and fresh lichen thalli by conventional methods has been difficult, because lichens contain high proteins, polysaccharides, and other complex compounds in their cell walls. Here we report a rapid, easy, and inexpensive protocol for extracting PCR-quality DNA from various lichen species. This method involves the following two steps: first, cell breakage using a beadbeater; and second, extraction, isolation, and precipitation of genomic DNA. The procedure requires approximately 10 mg of lichen thalli and can be completed within 20 min. The obtained DNAs were of sufficient quality and quantity to amplify the internal transcribed spacer region from the fungal and algal lichen components, as well as to sequence the amplified products. In addition, 26 different lichen taxa were tested, resulting in successful PCR products. The results of this study validated the experimental protocols, and clearly demonstrated the efficacy and value of our KCl extraction method applied in the fungal and algal samples.

• Journal of Microbiology and Biotechnology
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• 제20권12호
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• pp.1637-1646
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• 2010

The bacterium Deinococcus radiodurans is one of the most resistant organisms to ionizing radiation and other DNA-damaging agents. Although, at present, 30 Deinococcus species have been identified, the whole-genome sequences of most species remain unknown, with the exception of D. radiodurans (DRD), D. geothermalis, and D. deserti. In this study, comparative genomic hybridization (CGH) microarray analysis of three Deinococcus species, D. radiopugnans (DRP), D. proteolyticus (DPL), and D. radiophilus (DRPH), was performed using oligonucleotide arrays based on DRD. Approximately 28%, 14%, and 15% of 3,128 open reading frames (ORFs) of DRD were absent in the genomes of DRP, DPL, and DRPH, respectively. In addition, 162 DRD ORFs were absent in all three species. The absence of 17 randomly selected ORFs was confirmed by a Southern blot. Functional classification showed that the absent genes spanned a variety of functional categories: some genes involved in amino acid biosynthesis, cell envelope, cellular processes, central intermediary metabolism, and DNA metabolism were not present in any of the three deinococcal species tested. Finally, comparative genomic data showed that 120 genes were Deinococcus-specific, not the 230 reported previously. Specifically, ddrD, ddrO, and ddrH genes, previously identified as Deinococcus-specific, were not present in DRP, DPL, or DRPH, suggesting that only a portion of ddr genes are shared by all members of the genus Deinococcus.

• 한국균학회지
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• 제30권1호
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• pp.66-72
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• 2002

Phytophthora capsici 집단의 유전적 특성 분석용 DNA marker를 선발하고자 HindIII로 처리된 P. capsici 95CY3119 균주의 genomic DNA library를 임의로 cloning 시킨 후 southern blot 분석을 실시하여 선발된 clone들의 특성을 조사하였다. Probe로 사용하기 위해 선발된 clone 내에 삽입된 DNA 단편들은 HindIII로 처리된 P. capsici 95CY3119의 genomic DNA와 특이적으로 강하게 반응했다. 조사된 probe 중 PC9은 HindIII로 처리된 국내 P. capsici 균주들의 genomic DNA와 Southern 분석시 많은 밴드를 형성하였으며, 분리포장 단위별 균주들 간에는 물론 동일 포장 분리균주들 간에도 그 차이를 나타냈다. 그 밴드 양상을 기초로 집괴분석을 실시한 결과, 각 균주의 유전적 다양성이 잘 나타났다. Prove PC22는 다른 Phytophthora속과 Pythium속 균주들의 genomic DNA들과의 Southern hybridization에서는 반응을 나타내지 않았지만 특이적으로 P. capsici 균주들과는 다수의 밴드를 형성하였다. 이들 P. capsici 종특이적 DNA probe들은 추후 국내 및 전세계에 분포하는 P. capsici 집단의 유전적 다양성 분석 및 종동정에 유용한 marker로서 활용될 수 있을 것이다.