• 원예과학기술지
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• 제16권1호
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• pp.21-24
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• 1998

참외와 멜론의 다양성 분석을 위하여 RAPD 분석 최적조건과 군집분석하였다. 참외와 멜론계통의 DNA 추출은 0.5% SDS 방법이 가장 순수하고 많은 DNA를 얻을 수 있었으며 DNA 증폭시 최적 반응조건은 총 volum $15{\mu}L$ 중 DNA 10ng, Primer 270nM, dNTP $200{\mu}M$, dynazyme 0.3unit. 10x buffer $1.5{\mu}l$이였으며 나머지는 3차 증류수로 보충된다. PCR기기의 최적 setting은 DNA denaturation $94^{\circ}C$ 30초, primer annealing $39^{\circ}C$ 30초, DNA extension $72^{\circ}C$ 30초이며 최적 증폭 횟수는 40 cycle 이었다. 사용된 12개의 primer 만들어진 총 123개의 band 중 신뢰도가 높은 25개(20%)의 polymorphic band를 선발하여 이용하였으며 평균 polymorphic band 수는 2.1개로 나타났고, 그룹내 polymorphic band 수가 그룹간보다 적어 그룹내의 유전적변이가 적음을 보여주었다. 군집분석 결과 크게 참외와 멜론그룹으로 나뉘었고 멜론그룹은 다시 net melon 과 no-net melon으로 나뉘었으며 이러한 결과는 기존의 표현형질에 의한 분류와 일치하였다. 재래종 참외와 멜론 그룹은 8개의 marker에 의해 구분되었고 net melon 그룹과 no-net melon 그룹은 4개의 marker에 의해 구분되었다.

• 농업과학연구
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• 제26권2호
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• pp.39-48
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• 1999

본 연구는 축우 품종의 유전적 다형성을 분석하여 분자유전수준에서 축우의 품종간 판별과 유전적 유사성을 구명하고자 Holstein종과 한우, Charolais종 및 한우 교잡종의 혈액에서 DNA를 추출하여, PCR(polymerase chain reaction) 기법에 의한 RAPD(random amplified polymorphic DNAs)분석을 실시하고, 소 품종간의 유사도 지수를 추정한 후 UPGMA(unweighted pair-group method using average)방법으로 유사도를 추정하였으며 그 결과를 요약하면 다음과 같다. 1. 축우 품종들로부터 추출된 genomic DNA를 1.5% agarose gel에서 전기영동한 결과, 12.2kb 이상의 단일밴드로 나타났으며, genomic DNA의 순도는 DNA 흡광도 $A_{260}$과 $A_{280}$의 비율로 측정한 결과 그 비율은 1.75~2.10이었다. 2. RAPD기법에 의한 축우 품종의 유전적 다형성을 분석한 결과, 품종판별을 위한 가장 유용한 random primer는 5'-GAC CGC TTG T-3'인 것으로 판명되었다. 3. Primer 부착 온도에 따른 band 양상에 있어서 $33.0^{\circ}C$인 경우는 약 340bp에서 Holstein종과 Charolais종에서는 band가 출현되었으나, 한우와 한우 교잡종에서는 밴드가 나타나지 않았으며, $37.5^{\circ}C$의 경우에는 340bp에서 Holstein종과 한우에서는 밴드가 나타났으나, Charolais종과 한우 교잡종에서는 band가 출현되지 않아 이들 밴드의 유무가 소 품종판별을 위한 유용한 유전적 표지로서의 이용이 가능할 것으로 사료되었다. 4. 축우 품종의 유사도 지수에 있어서는 primer 부착온도를 $33.0^{\circ}C$와 $37.5^{\circ}C$을 종합하였을때 Holstein종과 한우간에는 0.666~0.777이었으며, Charolais종간에는 0.615~0.666이었고, 한우와 Charolais종간에는 0.400~0.461로 다소 낮은 수치를 보였으나, 한우와 한우 교잡종간에는 0.857~0.888로 대체로 높은 계수를 보였다. 5. 이상의 결과를 종합하여 보면 RAPD기법에 의하여 random primer 5'-GAC CGC TTG T-3'으로 축우 품종의 판별이 가능하며, UPGMA에 의한 축우 품종의 유사도는 Holstein종과 한우 및 Charolais종간 보다는 한우와 한우 교잡종 및 Charolais종간의 유전적 유사도가 높은 것으로 나타났다.

• 한국자원식물학회지
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• 제10권3호
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• pp.258-264
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• 1997

The introduction of molecular biology methodologies to plant improvement programs offers an invaluable opportunity for extensive germplasm characterization. We have applied the developed technique of random amplification of polymorphic DNA(RAPD)to the analysis of evaluating genetic diversity among Korean wild Codonopsis lanceolata. A total of 340 polymorpic hands were gernerated on agarose- and polyacrylamide-gel by 19 primers of abitrary sequence. grouped by cluster analysis using sample matching coefficients of similarity. Among of the samples. the minimum genetic distance value was obtained between sample no. 1(Girisan) and no. 2(Girisan), and the largest value between sample no. 11(Sulaksan) and no. 17(Sulaksan).In separate cluster dendrograms based on agareose - and polyacryamide-gel. some differences were observed; In the case of agarose gel,41 samples could be devided into 7 groups at below about 0.44 level of distance. However they were divided into 6 gourps at below about 0.40 level of distance in the case of polyacrylamide gel. These results showed that polymophic data in agrose were not grouped to wild plant selected from each mountainous district except for wild plants selected from Sulaksan and Chiaksan. We believe that polyacrylamide-RAPD is a superior method for detecting DNA polymorphism compared to agarose-RAPD method.

• Advances in Traditional Medicine
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• 제5권2호
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• pp.117-123
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• 2005

Dyers woad leaf (Daqingye) is a traditional Chinese medicine commonly used as anti-pyretic, anti-bacterial and anti-viral agent against infectious diseases. The Chinese Pharmacopoeia (2005) records that Dyers woad leaf should be derived from the leaves of Isatis indigotica Fort., but the leaves of Polygonum tinctorium Ait., Baphicacanthus cusia (Nees) Bremek. and Clerodendron cyrtophyllum Turcz. have also been used as substitutes of Dyers woad leaf in different regions of China. The leaf morphologies of these four species show a close resemblance, and based on their morphological appearance, it is difficult to identify them. Here, molecular genetic methods were developed as a target to identify different members of Dyers woad leaf. The 5S-rRNA spacer domain was amplified by polymerase chain reaction from genomic DNAs isolated from I. indigotica, P. tinctorium, B. cusia and C. cyrtophyllum, and the nucleotide sequences showed a great diversity. In addition, random amplification of polymorphic DNA analysis was also used to distinguish the members of Dyers woad leaf. These molecular methods could be used as a tool in authentic identification of Dyers woad leaf.

• 한국육종학회지
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• 제43권4호
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• pp.265-272
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• 2011

This study is to generate SCARs markers for identification of Perilla species. A SCAR is a genomic DNA fragment at a single genetically defined locus that is identified by PCR amplification using a pair of specific oligonucleotide primers. We derived SCARs by sequencing and cloning the both ends of the amplified products of RAPD markers. Sixteen sequence-specific primers were synthesized from eight RAPD markers, which were completely sequenced. We developed the species-specific SCAR markers which could be used successfully in detecting genetic variation in four Perilla species. These markers could be used to verify species-origins of various forms of Perilla germplasms.

• Plant Biotechnology Reports
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• 제4권3호
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• pp.237-242
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• 2010

Six parent and their 12 gamma ray-induced somatic flower colour mutants of garden rose were characterized to discriminate the mutants from their respective parents and understanding the genetic diversity using Random amplification of polymorphic DNA (RAPD) markers. Out of 20 primers screened, 14 primers yielded completely identical fragments patterns. The other 7 primers gave highly polymorphic banding patterns among the radiomutants. All the cultivars were identified by using only 7 primers. Moreover, individual mutants were also distinguished by unique RAPD marker bands. Based on the presence or absence of the 48 polymorphic bands, the genetic variations within and among the 18 cultivars were measured. Genetic distance between all 18 cultivars varied from 0.40 to 0.91, as revealed by Jaccard's coefficient matrix. A dendrogram was constructed based on the similarity matrix using the Neighbor Joining Tree method showed three main clusters. The present RAPD analysis can be used not only for estimating genetic diversity present in gamma ray-induced mutants but also for correct identification of mutant/new varieties for their legal protection under plant variety rights.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.130-130
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• 2003

Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

• 한국약용작물학회지
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• 제19권3호
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• pp.162-169
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• 2011

The rhizomes and herbal medicines originating from Acorus gramineus, A. calamus, A. tatarinowii, and A. gramineus var. pusilus, show significant similarity, and the correct identification of species is very difficult. Random Amplified Polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop a reliable method for identification of these four species. Several distinct SCAR markers were developed from species-specific RAPD amplicons for each species. Furthermore, a useful molecular marker was established for multiplex-PCR, in order to the four species could be distinguished concurrently. These markers allow efficient and rapid identification of closely-related Acorus species and will be useful for standardization of herbal medicines.

• Asian-Australasian Journal of Animal Sciences
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• 제17권9호
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• pp.1204-1209
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• 2004

Random amplification of polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out using DNA samples of 30 animals of Rathi cattle and 42 animals of Tharparkar cattle. Genomic DNA was isolated as per standard protocol and evaluated for its quality, purity and concentration. Twenty three random primers were screened out of which 15 primers yielded satisfactory amplifications and were used for further analysis. Average numbers of polymorphic fragments per primer were 7.07${\pm}$0.86 in Rathi and 6.80${\pm}$0.61 in Tharparkar cattle. The percentage of polymorphic bands in these two cattle breeds were 86 and 87%, respectively. Within breed genetic similarities for pooled over primers in the animals of Rathi and Tharparkar breeds were .577${\pm}$0.30 and 0.531${\pm}$0.02, respectively on the basis of band frequency (BF) and 0.645${\pm}$0.04 and 0.534${\pm}$0.04, respectively on the basis of band sharing (BS). Averages of between breed genetic similarities for pooled over primers were 0.97 and 0.92 according to BF and BS, respectively, which reflect higher degree of genetic similarity between Rathi and Tharparkar cattle breeds. Index of genetic distance based on BF and BS for pooled over primers was 0.030${\pm}$0.011 and 0.088${\pm}$0.031, respectively. Percentage of polymorphic bands and within-breed genetic similarities on the basis of band frequency (BF) and band sharing (BS) for pooled over primers revealed higher genetic similarity in Rathi than Tharparkar cattle population. High estimates of between breed genetic similarities for pooled over primers indicated that either Rathi is having decent from Tharparkar or both the cattle breeds are having common descent. Low value of Index of genetic distances between these two cattle breeds may be due to the fact that Rathi and Tharparkar cattle breeds are the native of Thar Desert in Northwest India. The results of between breed genetic distances also confirm the existence of high degree of genetic similarity between these two breeds of cattle.

• 한국동물위생학회지
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• 제41권3호
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• pp.149-155
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• 2018

Salmonella Enteritidis (S. Enteritidis) are found in animals, humans, and environment. In addition, S. Enteritidis draws attention to the public health concerns due to carriage of antibiotic resistance traits. For these reasons, the prevalence and antibiotic resistance patterns of S. Enteritidis are significant issues with regard to public health. To address this issues, a total of 24 strains of S. Enteritidis from 164 samples collected from several slaughterhouses in Gyeong-Nam province in order for antibiotic resistance profiles. Subsequently, we characterized the genotyping by random amplification polymorphic DNA (RAPD)-PCR. As a result, very high level of resistance to protein synthesis inhibition antibiotics and most isolates were susceptible to others. Six random primers were used for RAPD-PCR to reveal genotypes of S. Enteritidis isolates. One of the primer, P1245, generated 147 distinct RAPD-PCR fragments ranging from 400~3000 bp. The number of RAPD-PCR products ranged from 4 to 8 for this primer. The RAPD-PCR fragments could be placed these strains into 3 subgroups and 2 classes by UPGMA cluster analysis. Interestingly, several S. Enteritidis that isolated from different slaughterhouses showed same genotype. These results showed only limited genetic variation among the isolates, those were grouped into a few different patterns of antibiotic resistance.