• Title/Summary/Keyword: Randomly amplified polymorphic DNA (RAPD)

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Analysis of Lactobacillus casei and Mutant Strains by Polymerase Chain Reaction (Polymerase Chain Reaction에 의한 Lactobacillus casei 및 돌연변이 균주들의 비교 분석)

  • Nam, Jin-Sik;Lee, Jeong-Jun;Shin, Myeong-Su;Na, Seog-Hwan;Baek, Young-Jin;Yoo, Min
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.577-583
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    • 1994
  • To classify Lactobacillus casei strains on the basis of difference in their chromosomal DNA sequence, we have performed polymerase chain reactions on their chromosomal DNA by using random primers, and followed by analyzing randomly amplified polymorphic DNA fragments. We also developed a mini-preparative method to isolate PCR-grade chromosomal DNA from Lacto- bacillus casei strains within 3 hours. Based on RAPD pattems by polymerase chain reactions with degenerated random primers, 4 Lactobacillus casei strains and 2 mutant strains were successfully discriminated. Results were very sensitive, strain-specific and reproducible. It was also reliable. These results suggest that RAPD may be applied efficiently for the identification of several Lactoba- cillus casei strains.

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Studies on Genetic Stability of Micropropagated Plants and, Reintroduction in an Endemic and Endangered Taxon: Syzygium travancoricum Gamble (Myrtacae)

  • Ajith Anand
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.201-207
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    • 2003
  • Tissue culture techniques arguably are an important approach for ex situ conservation of rare and endangered plant species. However, there is utmost importance on maintaining the genetic integrity of the introduced plants especially in tree species. To examine the genetic integrity of the micropropagated plants, we randomly screened few hardened plants of Syzygium travancoricum, a critically endangered tree taxon, using Randomly Amplified Polymorphic DNA (RAPD) markers. Twenty-three random. primers were tried and twenty-five polymorphic loci were identified. The dendrogram based on the Unweighted Pair-Group Method Arithmetic Average and Nei's similarity index depicted about 97% homology between the mother plants and micropropagated plants. Further, an attempt was made to reintroduce the micropropagated plants in the wild. Over three hundred small trees could be successfully established.

Molecular Characterization of Selected Local and Exotic Cattle Using RAPD Marker

  • Khatun, M. Mahfuza;Hossain, Khondoker Moazzem;Rahman, S.M. Mahbubur
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.6
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    • pp.751-757
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    • 2012
  • In order to develop specific genetic markers and determine the genetic diversity of Bangladeshi native cattle (Pabna, Red Chittagong) and exotic breeds (Sahiwal), randomly amplified polymorphic DNA (RAPD) analysis was performed using 12 primers. Genomic DNA was extracted from 20 cattle (local and exotic) blood samples and extracted DNA was observed by gel electrophoresis. Among the random primers three were matched and found to be polymorphic. Genetic relations between cattle's were determined by RAPD polymorphisms from a total of 66.67%. Statistical analysis of the data, estimating the genetic distances between cattle and sketching the cluster trees were estimated by using MEGA 5.05 software. Comparatively highest genetic distance (0.834) was found between RCC-82 and SL-623. The lowest genetic distance (0.031) was observed between M-1222 and M-5730. The genetic diversity of Red Chittagong and Sahiwal cattle was relatively higher for a prescribed breed. Adequate diversity in performance and adaptability can be exploited from the study results for actual improvement accruing to conservation and development of indigenous cattle resources.

Development of Sequence Characterized Amplified Region Markers for Cultivar Identification in Persimmon (감 품종 판별용 SCAR 마커 개발)

  • Cho, Kang Hee;Cho, Kwang-Sik;Han, Jeom Hwa;Kim, Hyun Ran;Shin, Il Sheob;Kim, Se Hee;Chun, Jae An;Hwang, Hae-Sung
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.798-806
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    • 2013
  • Precise, fast, and cost-effective identification of crop cultivars is essential for plant breeder's rights. Traditional methods for identification of persimmon cultivars are based on the evaluation of sets of morphological characteristics. However, it is difficult to distinguish closely related cultivars using only morphological traits. This study was conducted to develop DNA markers for identification of the 32 persimmon cultivars in Korea and Japan. A total of 309 randomly amplified polymorphic DNA (RAPD) markers were identified using 40 different random primers. Various number of polymorphic bands ranged from 4 (OPP-08) to 14 (UBC159) were detected with an average of 7.7. Resulting 57 RAPD fragments were selected, and their sequences were determined for developing sequence characterized amplified region (SCAR) markers. As a result, 15 of 57 RAPD fragments were successfully converted to SCAR markers. Single polymorphic bands of the same size as or smaller than the RAPD fragments were amplified depending on SCAR markers. Among these markers, a combination of eight SCAR markers (PS225_200, PSN05_420, PSF13_523, PSN11_540, PS372_567, PS485_569, PSP08_635, and PS631_735) provided sufficient polymorphisms to identify 32 persimmon cultivars. These newly developed markers will be a fast and reliable tool to identify persimmon cultivars.

RAPD-SCAR Markers Linked to Medium-Leaf Zoysiagrass Ecotypes (한국잔디 중지 변이개체와 연관된 RAPD-SCAR 마커)

  • Chung, Sung Jin;Park, Su Jeong;Kim, Hun Joong;Yang, Geun-Mo;Choi, Joon-Soo;Oh, Chan-Jin;Jang, Deok-Hwan;Song, In-Ja;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.2 no.2
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    • pp.191-197
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    • 2013
  • Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

Comparison of RAPD Profiles and Phenotypical Characters of Streptococcal Strains (연쇄상구균의 표현형적 특성과 RAPD profiles 비교)

  • Song, Jin-Gyeong;Kim, Jong-Hun;Kim, Eun-Hui
    • Journal of fish pathology
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    • v.16 no.1
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    • pp.51-59
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    • 2003
  • Streptococcal infection is one of the most serious disease of cultured olive flounder, Paralychthys olivaceus in Korea and caused by more than one species. However, there has been considerable confusions about the taxonomic position of the fish pathogenic streptococci. In this study, We performed the randomly amplified polymorphic DNA(RAPD) pattern analysis to evaluate the possible classification in 8 streptococci isolated from diseased olive flounder and reference strains based on their DNA structure. RAPD PCR with DNA solution prepared by simple boiling and 10-mer random primer was appeared to be a good tool for discrimination of different streptococcal strains. Phenotypical characters by simple biological test and API 20 Strep corresponded well to the specific profiles of RAPD in streptococcal isolates of this study. Therefore, the RAPD profile was considered as one of differential characters to discriminate the streptococcal isolates from diseased olive flounder.

Discrimination of Bacillus anthracis from Bacillus cereus Group Using KHT5 Marker (KHT5 마커를 사용한 Bacillus cereus 그룹에서 Bacillus anthracis의 구별)

  • 김형태;김성주;채영규
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.40-44
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    • 2003
  • Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. In order to develop a DNA marker specific for Bacillus anthracis and to discriminate this species from Bacillus cereus group, we applied the randomly amplified polymorphic DNA (RAPD)-PCR technique to a collection of 29 strains of the genus Bacillus, including 22 species of the B. cereus group. A 709-bp RAPD marker (KHT5) specific for B. anthracis was obtained from B. anthracis BAK. The PCR product of internal primer set from the KHT5 fragment distinguished B. anthracis from the other species of the B. cereus group.

Analysis of Genetic Relatedness by Random Amplified Polymorphic DNA (RAPD) in Pecan Taxa (RAPD를 이용한 Pecan 품종의 유전적 관계 분석)

  • 신동영;김회택;박종인;노일섭
    • Korean Journal of Plant Resources
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    • v.13 no.1
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    • pp.1-10
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    • 2000
  • Pecan is deciduous tree and belongs to the Julandaceae family. Pecan is an economically important as a nut and timber crop. Heterozygosity is expected to be high for typically cross-pollinated. Yet little is known about the nature of genetic variation within this species. In addition, the pedigree of many pecan cultivars remains unknown or is questionable. In this study, the phylogenetic relationships between 22 pecan cultivars and its analyzed by RAPD (randomly amplified polymorphic DNA). PCR Amplification used 40 randomly selected oligoes as primers. Based on their genetic similarities derived from the RAPD data, the 22 pecan cultivars were classified into different five groups in agarose gel. The 22 pecan cultivars were classified into five sectional groups by UPGMA clustering analysis, too. C. flacra and Black walnut showed the 0.9 of similarity index and Farley, Pawnee showed the 0.85 of similarity index. The 22 pecan cultivars were classified into different five groups by analysis of the 4% polyacrylamide gel fraction. (Group I : 1, 2, 3, 4, 13, 16, 17, 20, 21 Group II : 14,18 GroupIII : 6,12 GroupIV : 5, 11, 15, 19, 22 CroupV : 7, 8, 9, 10) Group V show the 1.0 of similarity index and Farley, Sturya, Clarke, Pawnee show the 0.98 of similarity index and Kiowa, Schley show the 0.92 of similarity index. Results from this study indicated that RAPD can be used to establish the genetic relationships among the 22 pecan cultivars. Similarity coefficients generally agreed with what would be predicted in cultivars with known pedigrees, and we could accurately construct relationships among cultivars. In addition, we have shown that RAPD provides useful information on the origin of unknown cultivars.

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SCAR markers were developed to identify zoysiagrass mutants exhibiting fine leaf characteristics (세엽 한국들잔디 변이체 식별을 위한 SCAR 마커 개발)

  • Chung, Sung Jin;Park, Su Jeong;Choi, Young In;Kim, In-Kyung;Lee, Ka-Yeon;Kim, Hun-Joong;Lee, Geung-Joo
    • Korean Journal of Agricultural Science
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    • v.40 no.2
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    • pp.115-121
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    • 2013
  • Polymorphic bands of two fine-leaf zoysiagrass mutants (CNU 70-1, CNU 70-2) induced via a gamma-ray irradiation on seeds of Zoysia japonica were obtained by using randomly amplified polymorphic DNA (RAPD) primers. The genotype-specific fragments were then converted into PCR-based sequence characterized amplified region (SCAR) markers, which are now amenable to detecting them among other zoysiagrass species widely noticeable in Korea. The CNU 70-1-specific primer set amplified about 900 bp successfully, while the CNU 70-6 marker produced the expected 1,500 bp band, by which those markers were nominated by CNU 70-1_900 and CNU 70-6_1500 SCARs, respectively. The developed SCAR markers can be an applicable tool in sod industry where illegal appropriation hampers breeder's right and profits due to the turfgrass plant vegetatively propagating.

Randomly Amplified Polymorphic DNA Analyses of Pestalotiopsis theae Isolated from Sweet Persimon (재배되는 단감나무로 부터 분리한 Pestalotiopsis theae의 RAPD 기법을 이용한 유전특성의 비교분석)

  • Lee, Youn-Su;Woo, Su-Jin;Choi, Hei-Sun;Kim, Kyoung-Su;Kang, Won-Hee;Kim, Myoung-Jo;Shim, Jae-Ouk;Chang, Tae-Hyun;Lim, Tae-Heon
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.365-372
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    • 1998
  • In this study, we evaluated the genetic relationships of fourty seven Pestalotiopsis theae isolates collected from diseased sweet persimon in various places in southern part of Korea using RAPD (Randomly Amplified Polymorphic DNAs) method. As a result of the amplification, eight primers showed total of 86 bands ranging from 0.3 Kb to 3.2 Kb. Among those 86 bands, 84 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Similarities among the compared isolates ranged from below 60% to more than 95%. Most of the compared isolates showed $50{\sim}80%$ similarities. The number of isolate pairs which showed more than 80% similarity were 248. The number of isolate pairs which showed $50{\sim}80%$ similarity were 789, and the number of isolate pairs which showed below 50% similarity were 21. Isolate SP-21 (No.9) showed below 50% similarity with all the isolates compared. At 50% similarity level, all the isolates compared, except isolate SP-21 (No.9), were included in one big group. At 65% similarity level, all the isolates compared, except isolate SP-21 (No.9), were divided into three different groups. At 75% similarity level, all the isolates compared, except isolates SP-47 (No. 23) and SP-21 (No.9), were divided into six different groups.

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