• Journal of Microbiology
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• 제44권1호
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• pp.106-112
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• 2006

In this study, the genetic diversities of multi-resistant Salmonella typhimurium (ST) isolates were analyzed via the application of both pulsed field gel electrophoresis (PFGE) and Polymerase chain reaction (PCR) analysis methods, using 6 kinds of primers (REP, ERIC, SERE, BOX, P-1254 and OPB-17). And their discriminative abilities (DA) were also compared in order to determine the most effective and reliable analysis method. 118 S. typhimurium isolates, cultured from diverse animals and human patients in Korea beginning in 1993, were analyzed and subjected to a comparison of Simpson's index of diversity (SID), using both PFGE and PCR methods. PFGE by XbaI enzyme digestion allowed for discrimination into 9 pulsotypes, with high SID values (0.991) on the genomic DNA level. This shows that PFGE is a very discriminative genotypic tool, and also that multiple clones of S. typhimurium isolates had existed in domestic animals and humans in Korea since 1993. However, we could ultimately not to trace the definitive sources or animal reservoirs of specific S. typhimurium isolates examined in this study. Depending on the SID values, the combined method (7 kinds of method) was found to be the most discriminative method, followed by (in order) SERE-PCR, REP-PCR, ERIC-PCR, PFGE & OPB-17 (RAPD), P-1254 (RAPD), and BOX-PCR at the $80\%$ clone cut-off value. This finding suggests that the REP-PCR method (which utilizes 4 primer types) may be an alternative tool to PFGE for the genotyping of S. typhimurium isolates, with comparable cost, time, and labor requirement. The establishment of a highly reliable and discriminatory method for epidemiologic analysis is considered necessary in order for researchers to trace the sources of specific pathogens and, consequently, to control and prevent the spread of epidemic S. typhimurium isolates to humans.

• 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.

• Fisheries and Aquatic Sciences
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• 제5권3호
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• pp.156-164
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• 2002

Enteromorpha prolifera of the isomorphic diploid sporophyte and the haploid gametophyte generations inhabit rocks, tidal flats and tidal pools in the middle parts of intertidal zones. In this experiment, their thalli were observed by bare eyes from October and experienced $74\pm16.5cm$ maximum growth the following March and April. The rate of occurrence of the thalli per month was highest in March, while their biomass peaked at $1,464\pm41.5 g/m^2$ in Jangheung in April. Genetic similarity was investigated samples of E. prolifera collected from Muan, Wando, Jangheung, Yosu and Jinhae, at the south coast of Korea. Random amplified polymorphic DNA (RAPD) markers were used. For the RAPD analysis, 3 ng of the DNA extracted from the thalli using he phenol/chloroform method was amplified by PCR with a 25 {\mu}L$ reaction solution, arbitrary primers and 36 cycles. Among the 60 primers used, 31 yielded products, most of which showed diverse electrophoresis patterns. Similarities among the groups compared ranged from 0.37 to 0.58. We conclude that the use of RAPD analysis is appropriate to characterize the genetic variability of this commercial species along its geographical distribution.

• 한국작물학회지
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• 제46권4호
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• pp.341-343
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• 2001

Molecular markers have become fundamental tools for crop genome study. The objective of this study was to construct a genetic linkage map for cowpea with PCR-based molecular markers. Five hundred and twenty random RAPD primers were screened for parental polymorphism. Ninety RAPD markers from sixty primers was segregated in 75 F2 mapping population derived from the cross of local cultivars GSC01 and GSC02. 70 RAPD markers were found to be genetically linked and formed 11 linkage groups. Linkage map spanned 474.1 cM across all 11 linkage groups. There are six linkage groups of 40 cM or more, and five smaller linkage groups range from 4.9 to 24.8 cM. The average linkage distance between pairs of markers among all linkage groups was 6.87 cM. The number of markers per linkage group ranged from 2 to 32. The longest group 1 spans 190.6 cM, while the length of shortest group 11 is 4.9 cM. This map is further needed to be saturated with the various markers such as RFLP, AFLP, SSR and more various populations and primers. In addition, morphological markers and biochemical markers should be united to construct a comprehensive linkage map.

• The Plant Pathology Journal
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• 제23권2호
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• pp.45-50
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• 2007

Genetic diversity among selected isolates of Macrophomina phaseolina, a causal agent of charcoal rot (stalk rot) disease in sorghum was studied using PCR-RAPD markers. A set of ten isolates, from ten different rabi sorghum genotypes representing two traditional sorghum growing situations viz., Dharwad- a transitional high rainfall region and Bijapur- a semi-arid low rainfall region in South India. From a set of 40 random primers tested, amplicon profiles of 15 were reproducible. A total of 149 amplicon levels, with an average of 9.9 bands per primer, were available for analysis, of which 148 were polymorphic (99.3%). It was possible to discriminate all the isolates with any of the 15 primers employed. UPGMA clustering of data indicated that the isolates shared varied levels of genetic similarity within a range of 0.14 to 0.72 similarity coefficient index and it was suggestive that grouping of isolates was not related to sampling location in anyway. A high level of genetic heterogeneity of 0.28 was recorded among the isolates.

• 아시안잔디학회지
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• 제13권3호
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• pp.153-158
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• 1999

'Konwoo', bermudagrass[Cynodon dactylon (L) Pers.] (Patent registration no. 2000-724), a vegetative cultivar, was developed by the Dept. of Horticultural Science, Konkuk University, Seoul. 'Konwoo' was selected in 1997 among 20 lines collected from Korea, Japan, Tiwan, China, U. S. A, and Australia. 'Konwoo' morphologically similar to Tifway 419 was selected due to the erect type, short leaf length($1.3\pm$0.3cm), fine leaf($2.0\pm$0.5mm), rapid establishment and recoverage, many stolon number and high shoot density. When 'Konwoo' was compared to the four other bermudagrass lines at the DNA level using 54 PCR primers, it had the specific bands with primer No. 102, 275, 280, 295, 300, 739 by RAPD analysis.

• Asian-Australasian Journal of Animal Sciences
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• 제15권8호
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• pp.1071-1075
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• 2002

In order to identify and develop the specific DNA marker for the identification of Hanwoo (Korean Cattle) from other breeds, a specific DNA marker of 519 bp was identified and sequenced from polymorphic analysis using RAPD-PCR for 6 cattle breeds. Two different repetitive sequences, $(AAC)_5$ and $(GAAGA)_2$, were selected and designed to use specific probe to develop a DNA marker for Hanwoo specific. When the $(AAC)_5$ probe was applied, the 10 kb specific DNA marker showed in the DNA fingerprinting from 237 of 281 Hanwoo individuals. This novel Hanwoo specific DNA probe is useful to perform the marker-assisted selection for screening Hanwoo purity as an unique genetic source.

• Food Science and Biotechnology
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• 제14권6호
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• pp.875-877
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• 2005

Presence of Enterobacter sakazakii, occasional pathogen of powdered infant formula causing rare, but life-threatening diseases such as neonatal meningitis, bacteremia, necrotizing enterocolitis, and necrotizing meningoencephalitis after ingestion was examined in 45 powdered infant formula products manufactured in Korea using chromogenic Druggan-Forsythe-Iversen (DFI) medium, and isolates were identified with API 20E. Ent. sakazakii was isolated from three products. Ent. sakazakii isolates were genotyped by RAPD-PCR using two random primers, and their banding patterns were compared.

• 생명과학회지
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• 제11권1호
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• pp.65-67
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• 2001

In order to develop a rapid and simple method for testing the purity of radish hybrid seeds using a procedure based on the PCR(Polymerase chain reaction), eighty random primers were screened with the genomic DNA extracted from five day old seedlings of inbred parent lines and their F1 hybrids. Two primers, HRM-02 (5'-GAGACCAGAC-3') and HRM-19(5'-TGAGGCGTGT-3'), generate reproducible unique PCR patterns which can identify each parent lines as well as their hybrids. In actual test of randomly selected hybrid seeds using the two marker primers, the purity tested by one primer was exactly same as that of other primer. It suggests that one marker primer selected in this experiment is enough for the purity test of radish hybrid seeds. We demonstrates the use of RAPD(randomly amplified polymorphic DNAs) markers to identify each of inbred parent lines and hybrids by rapid and simple method.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.85.1-85.1
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• 2003

Previously, we found the operon random primer (OP-5A) that is characteristic the genus Panax by randomly amplified polymorphic DNA (RAPD) analysis. However, OP-5A primer is limited to apply on the differentiation of only crude herbal plants. To construct more sensitive and unique primers on the genus Panax, ginseng-specific DNA profile (350 bp) that was amplified by OP-5A primer were inserted in a plasmid vector in the TA cloning method and sequenced. We designed the PCR primers (Forward: 5＂-AGGGGTCTTGCTAT AGCGGAAC-3＂, Reverse: 5＂-AGTCTTAATTTCATATTTTCGTATG-3＂) and identified the unique ginseng band (350 bp) in commercial granule products including ginseng extracts as well as crude ginseng plants by nascent PCR.(omitted)