• Title/Summary/Keyword: Random amplified polymorphic DNAs

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Genetic Similarity Frequency and DNA Polymorphism between Common Carp and Israeli Carp Using Polymerase Chain Reaction-Random Amplified Polymorphic DNAs

  • Yoon, Jong-Man;Park, Min-Soon;Kim, Young-Gill
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.334-335
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    • 2001
  • Common carp (Cyprinus carpio) and Israeli carp(C. carpio) samples were obtained from a aquaculture facility in the Kunsan National University, Korea. Genomic DNA was isolated from the common carp and Israeli carp representing genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA as arbitrary primers. There were observed a total of 90 species-specific genetic markers within Israeli carp. On average, each random RAPD primer produced amplified 7.9 products from 1 to 17 bands. An average genetic similarity within Israeli carp showed -.60$\pm$0.05. The average level of bandsharing was some 0.57$\pm$0.03 between common carp and Israeli carp. Accordingly, two carp species were genetically a little distant. The electrophoretic analysis of PCR-RAPD proudcts showed high levels of variation between two fish species. The RAPD polymorphism generated by primer may be used as a genetic marker for species or lines identification in important aquacultural carp.

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SCAR Marker Linked with A1 Mating Type Locus in Phytophthora infestans

  • Zhang Xuan-Zhe;Seo Hyo-Won;Ahn Won-Gyeong;Kim Byung-Sup
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.724-730
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    • 2006
  • A sequence characterized amplified region (SCAR) marker, which was tightly linked with the A1 mating type locus in Phytophthora infestans, was developed. During the random amplified polymorphic DNA-based phylogenic studies of 33 isolates of P infestans collected from year 2002 to 2004, we found an A1 mating type-specific DNA fragment. This 573-bp DNA fragment was generated only in the genomic DNA of the A1 mating types, when OPC-5 primer was used. Based on the specific DNA sequence, we designed the primer sets for generating the A1 mating type-specific 569-bp DNA fragment. When 33 genomic DNAs of P. infestans were subjected to PCR amplification using different primer combinations, the A1 mating type-specific DNA was amplified, when LB-1F and LB-2R primers were used. The specific 569-bp DNA fragment was generated only from all 18 A1 strains, but not from 15 A2 mating type strains. These results corresponded to the mating type discriminating bioassay of 33 isolates of P. infestans. Therefore, the primer combination of LB-1F/LB2R was chosen as a SCAR marker. Overall, this study indicates that the SCAR marker could be developed into a useful tool for mating type determination of P. infestans.

Species identification of Dyers woad leaf by DNA sequence of 5S-rRNA spacer domain and random amplified polymorphic DNA (RAPD) analysis

  • Zhao, K.J.;Dong, T.T.X.;Wong, Y.C.;Tu, P.F.;Tsim, K.W.K.
    • Advances in Traditional Medicine
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    • v.5 no.2
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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.

Genetic Differences of Two Wild Shortnecked Clam(Ruditapes philippinarum) Populations from the Yellow Sea Analysed by Random Amplified Polymorphic DNAs-Polymerase Chain Reaction

  • Yoon, Jong-Man;Kim, Yong-Ho
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2003.05a
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    • pp.229-230
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    • 2003
  • Shortnecked clam is a commercially important mollusks species, which is distributed all over the Yellow Sea. Consequent of the rapid increase in seed production, there is a need to understand the genetic composition of wild shortnecked clam populations in order to evaluate exactly the latent genetic effects induced by seed production operations. Many genetic and molecular researches were made because RAPD-PCR is a simple and rapid method for determining genetic diversity and similarity in various organisms with the advantage that no prior knowledge of the genome under research is needed (Fischer et at., 2000). (omitted)

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Variations of RAPD and Chemical Composition of Capsositiphon fulvescens Culturing in Korea

  • Sun, Sangmi;Chung, Gyuhwa
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.169-170
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    • 2000
  • The green marine algae, Capsosiphon fulvescens has been cultivated in south coast of southern Korea for many years on a commercial scale. This species is very popular in Korean as a food supplement because of its attractive flavor and flexcible taste. It is, therefore, necessary to isolate and utilize qualified germplasms for mass production of this economic seaweed. Several reports have been published on phycological applications of RAPDs including the characterization of interspecific genetic variation, the identification of isolates and hybrids, and the study of phylogenetic relationships. However few authors have used RAPDs to assess the genetic variability among populations of a seaweed species(van Oppen et al., 1994; Alberto et al., 1997). The present study was undertaken for characterizing the identities of Capsosiphon fulvescens populations cultivating in Korea through the analysis of PCR based random amplified polymorphic DNAs (Welsh and MacClelland, 1990; Willams et al., 1990) and chemical composition aimed to isolate the useful strains for aquaculture. (omitted)

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In Vitro Regeneration of Lycium chinense Miller and Detection of Silent Somaclones with RAPD Polymorphisms

  • Ahn, In-Suk;Park, Young-Goo;Shin, Dong-Ill;Sul, Ill-Whan
    • Journal of Plant Biotechnology
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    • v.6 no.3
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    • pp.157-163
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    • 2004
  • An efficient system for the regeneration of adventitious shoots from in vitro cultured leaf sections of Lycium chinense Miller was developed and silent somaclones from the regenerants detected with RAPD method. Among the eight media tested (B5, SH, N&N, 1/2MS, MS, 3/2MS, GD and WPM), and four cytokinins (BA, kinetin, 2ip and zeatin) with different concentrations (1, 5, 10, 20, 30 and 40 $\mu{M}$), 1/2 MS medium supplemented with 20 and 30 $\mu{M}$ zeatin showed the best regeneration frequency (100% and 93.7%) and higher average number of shoots (9.0 and 9.4). All regenerants easily elongated after subculturing on 1/4MS without growth stimulants and produced spontaneous adventitious roots from their basal parts. With phenotypically normal 40 regenerants, RAPD analysis with 15 different random primers was performed to examine the cryptic somaclonal variants. No substantial differences in banding patterns were found in the amplified polymorphic DNAs implying no DNA changes during dedifferentiation into adventitious shoots. However, one (OPF-4) of the 15 primers detected silent somaclonal variation in one regenerant in which two different polymorphic bands did not appear when compared with the rest regenerants. The results indicate that regenerantion via intervening callus phase can be used to establish true-to-type planting stocks for homogeneous population.

Genetic Diversity Analysis of the Cheju Horse Using Random Amplified Polymorphic DNAs (PCR-RAPD를 이용한 제주말의 유전적 다양성분석)

  • Cho, Byung-Wook;Lee, Kil-Wang
    • Journal of Life Science
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    • v.14 no.3
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    • pp.521-524
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    • 2004
  • This experiment was carried out to analyze genetic characteristics and to develop the breed specific DNA marker for Cheju-native horse. If this marker contains high repetitive sequences, it is possible to convert a RAPD marker of interest into a single-locus PCR marker called a sequence characterized amplified region(SCAR). Twenty six Cheju-native horse and Fifty thoroughbred genomic DNA were pooled and PCR. were accomplished using 800 random primers. Comparing the pooled DNA from Cheju-native horse and thoroughbred, we found 9 primers which identified markers present in the pooled DNA from breed but absent in the other breed. Among 9 random primers, 6 primers were thoroughbred specific and 3 primers were Cheju-native horse specific. Testing individual horse revealed that 5 marker showed the similar band pattern between Cheju-native horse and Thoroughbred. However, 4 marker were wholly absent in breed while present in the other breed. UBC $126_{3500bp}$, UBC $162_{500bp}$, and UBC $244_{1200bp}$ was detected only Thoroughbred and UBC $562_{560bp}$was detected Cheju-native horse, respectively. After determining of the cloned breed-specific fragment sequence, we designed the SCAR-primers and carried out PCR. Compared to random primer, RAPD-SCAR primer didn't show significantly higher specific band. However, RAPD analysis is useful for genetic characterization of Cheju-native horse.

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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Absence of DNA Polymorphisms in Myzus persicae (Homoptera: Aphididae) in Relation to their Host Plants (기주식물 종류에 따른 복숭아혹진딧물(Myzus persicae)의 DNA Polymorphism 비교)

  • H. J. Kim;K. S. Boo;K. H. Cho
    • Korean journal of applied entomology
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    • v.35 no.3
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    • pp.209-215
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    • 1996
  • DNA polymorphisms were analyzed for 8 clones of the green peach aphid, Myzus persicae Sulzer, by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). The insect has different host preferences and was even classified into two different species, M. persicae Sulzer and Myzus nicotinae Blackman by their morphological characters, but this point is still in arguement. To identify the differences between two types of the green peach aphid by RAPD-PCR, the template DNA was extracted from 4 clones each of tobacco-feeding and non-tobacco-feeding forms and one hundred primers of 10-nucleotideslong were tested in PCR. The amplified DNAs were analyzed by agarose gel electrophoresis. Eighty-three primers gave amplified DNA fragments with 1 to 22 in number and 500 to 20,000 base pairs in length, but no amplification was observed in the other 17 primers. The average number of fragment per each amplification was about 13. In the case of 82 out of 83 random primers, band patterns of amplified DNA were identical among 8 clones, even though some differences were noticed in the intensity of specific bands. Polymorphism was detected by only one primer within the tobacco-feeding forms, but not between the two host types. The results did not detect any relationship between RAPD polymorphism and their host preference.

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RAPD Identification of Genetic Variation in Ulvales Seaweed (RAPD기법을 이용한 갈파래목 해조류의 유전 변이 분석)

  • CHO Yong-Chul;PARK Ji Won;JIN Hyung-Joo;NAM Bo-Hye;SOHN Chul Hyun;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.388-392
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    • 1997
  • The random amplified polymorphic DNAs (RAPD) technique was used to characterize seven isolates of the green seaweed Ulvales collected from Songjeng, Haeundae, Jumunjin, Dadaepo and Wando in Korea. Total DNA was extracted by the LiCl extraction method from thalli of green seaweed. The extracted DNA (3 ng) in $25{\mu}\ell$ reaction volume was amplified by 45 cycles of the polymerase chain reaction with arbitrary primers. Thirty-four primers resulted in 1227 PCR products ranged 240 bp to 1.5 kb of both conserved and polymorphic bands. Genetic similarities of the seven isolates calculated by Jaccard's equation were ranged from $7\%\;to\;36\%$. Monostroma nitidum (Wando) was shown to be most distantly related with the others based on genetic similarity and did not produce the amplified band of 630 bp, common in Ulvales using primer OPB-01 (CATCCCCTG).

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