• Title/Summary/Keyword: polymorphic

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PCR Based Detection of Phellinus linteus using Specific Primers Generated from Universal Rice Primer(URP) Derived PCR Polymorphic Band

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;Lee, Byoung-Moo;Cho, Soo-Muk;Kim, Ki-Tae;Seo, Geon-Sik;Go, Seung-Joo
    • Mycobiology
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    • v.30 no.4
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    • pp.202-207
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    • 2002
  • This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

Identification of New Microsatellite Markers in Panax ginseng

  • Kim, Joonki;Jo, Beom Ho;Lee, Kyoung Lyong;Yoon, Eui-Soo;Ryu, Gi Hyung;Chung, Ki Wha
    • Molecules and Cells
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    • v.24 no.1
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    • pp.60-68
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    • 2007
  • Microsatellites, also called simple sequence repeats (SSR), are very useful molecular genetic markers commonly used in crop breeding, species identification and linkage analysis. In the present study, we constructed a microsatellite-enriched genomic library of Panax ginseng, and identified 251 novel microsatellite sequences. Tri-nt repeat units were the most abundant (46.6%), followed by di-nt repeats (35.5%). The $(AG)_n$ motif was most common (23.1%), followed by the $(AAC)_n$ motif (22.3%). From the genotyping of 94 microsatellites using marker-specific primer sets, we identified 11 intraspecific polymorphic markers as well as 14 possible interspecific polymorphic markers differing between P. ginseng and P. quinquefolius. The exact allele structures of the polymorphic markers were determined and the alleles were named. This study represents the first report of the bulk isolation of microsatellites by screening a microsatellite-enriched genomic library in P. ginseng. The microsatellite markers could be useful for linkage analysis, genetic breeding and authentication of Panax species.

Differentiation of Salmonella typhimurium from Gram-negative Intestinal Microbes by Randomly Amplified Polymorphic DNA (RAPD) Fingerprinting

  • Jin, Un-Ho;Chung, Tae-Wook;Kim, June-Ki;Nam, Kyung-Soo;Ha, Sang-Do;Kim, Cheorl-Ho
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.8-10
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    • 2000
  • In order to rapidly identify and differentiate Salmonella typhimurium from the intestinal gram-negative bacteria, randomly amplified polymorphic DNA (RAPD) fingerprinting of Salmonella typhimurium was carried out using random primers designated OPA-13 (5'-CAGCACCCAC-3'), OPB-10 (5'-CGTCTGGGAC-3'), OPB-18 (5'-CCACAGCAGT-3'), and OPJ-10 (5'-AAGCCCGAGG-3'), and its patterns compared with 6 representive intestinal, gram-negative bacterial strains, Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, Escherichia coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., which are often found in foods. S. typhimurium had unique and distinct fingerprinting patterns. RAPD fingerprinting is thus concluded to be a rapid and sensitive method for the identification of S. typhimurium compared to conventional culturing procedures or immunoassays.

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Thoracoscopic Left Cardiac Sympathetic Denervation for a Patient with Catecholaminergic Polymorphic Ventricular Tachycardia and Recurrent Implantable Cardioverter-Defibrillator Shocks

  • Yu, Woo-Sik;Kim, Tae-Hoon;Suh, Jee Won;Song, Seunghwan;Lee, Chang Young;Joung, Boyoung
    • Journal of Chest Surgery
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    • v.48 no.3
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    • pp.220-224
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    • 2015
  • A patient presented with loss of consciousness and conversion. During an exercise test, catecholaminergic polymorphic ventricular tachycardia (CPVT) resulted in cardiac arrest. He started taking medication (a beta-blocker and flecainide) and an implantable cardioverter defibrillator (ICD) was inserted, but the ventricular tachycardia did not resolve. Left cardiac sympathetic denervation (LCSD) was then performed under general anesthesia, and the patient was discharged on the second postoperative day without complications. One month after the operation, no shock had been administered by the ICD, and an exercise stress test did not induce ventricular tachycardia. Although beta- blockers are the gold standard of therapy in patients with CPVT, thoracoscopic LCSD is safe and can be an effective alternative treatment option for patients with intractable CPVT.

Genetic Diversity and DNA Polymorphism in Platycodon grandiflorum DC. Collected from East-Asian Area

  • Park, Chun-Geun;Yan, Zhi-Yi;Lee, Sang-Chul;Shon, Tae-Kwon;Park, Hee-Woon;Jin, Dong-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.2
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    • pp.115-120
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    • 2005
  • Broadening the genetic base of Platycodon grandiflorum DC. cultivar to sustain improvement requires assessment of genetic diversity available in P. grandiflorum DC.. The objective of this study was to analyze the genetic variation, genetic relationship among 48 samples collected from East-Asian Area by means of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) markers. From the 18 primers tested, produced total 211 bands with an average of 11.7 bands per primer and obtained 103 polymorphic band with an average of 5.7 bands per primer,s revealed relatively high percentage of polymorphic bands (48.8%). The genetic similarities calculated from RAPD data varied from 0.688 to 0.994 and were clustered to six major groups on a criterion of 0.78 similarity coefficient. The present study has revealed the significant genetic similarity among the samples tested. The analysis of genetic relationships in P. grandiflorum using RAPD-PCR banding data can be useful for the breed improvement.

Screening of SHOX gene sequence variants in Saudi Arabian children with idiopathic short stature

  • Alharthi, Abdulla A.;El-Hallous, Ehab I.;Talaat, Iman M.;Alghamdi, Hamed A.;Almalki, Matar I.;Gaber, Ahmed
    • Clinical and Experimental Pediatrics
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    • v.60 no.10
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    • pp.327-332
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    • 2017
  • Purpose: Short stature affects approximately 2%-3% of children, representing one of the most frequent disorders for which clinical attention is sought during childhood. Despite assumed genetic heterogeneity, mutations or deletions in the short stature homeobox-containing gene (SHOX ) are frequently detected in subjects with short stature. Idiopathic short stature (ISS) refers to patients with short stature for various unknown reasons. The goal of this study was to screen all the exons of SHOX to identify related mutations. Methods: We screened all the exons of SHOX for mutations analysis in 105 ISS children patients (57 girls and 48 boys) living in Taif governorate, KSA using a direct DNA sequencing method. Height, arm span, and sitting height were recorded, and subischial leg length was calculated. Results: A total of 30 of 105 ISS patients (28%) contained six polymorphic variants in exons 1, 2, 4, and 6. One mutation was found in the DNA domain binding region of exon 4. Three of these polymorphic variants were novel, while the others were reported previously. There were no significant differences in anthropometric measures in ISS patients with and without identifiable polymorphic variants in SHOX. Conclusion: In Saudi Arabia ISS patients, rather than SHOX, it is possible that new genes are involved in longitudinal growth. Additional molecular analysis is required to diagnose and understand the etiology of this disease.

Evaluation of ISSR and RAPD Markers for the Detection of Genetic Diversity in Mulberry (Morus spp.)

  • Venkateswarlu, M.;Nath, B.Surendra;Saratchandra, B.;Urs, S.Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.207-215
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    • 2004
  • The present study was carried out to evaluate the ISSR and RAPD markers for their efficiency as genetic marker systems to establish the relationships between 18 mulberry genotypes. A total of 36 from 56 (64%) RAPD primers and 12 from 48 (25%) ISSR primers produced reproducible amplification patterns. A high proportion of polymorphic bands ranging from 44 to 91% was observed respectively with RAPD and ISSR markers. The average Resolving Power (Rp) of ISSR primers was higher than RAPD primers. The ISSR primers, UBC 825, 868 and 873, and RAPD primers, UBC 712, 720 and 729, possessed the highest Rp values and could in each instance distinguish all the 18 genotypes. Similarity matrix values were estimated based on Jaccards coefficient, considering 109 polymorphic ISSR and 212 polymorphic RAPD bands and two dendrograms were constructed. The dendrograms obtained with ISSR and RAPD markers distinguished the eight exotic genotypes from the ten indigenous (Indian) genotypes. A significant correlation value (r=0.959; p=0.001) for the cophenetic matrix between the RAPD and ISSR matrices was observed. The results indicated that the ISSR and RAPD markers could assist in the differentiation of genotypes and permit the determination of genetic distances that might be exploited by mulberry breeders in improvement programs.

Specific Detection of Erwinia carotovora subsp. carotovora by DNA Probe Selected from PCR Polymorphic Bands (PCR다형성 밴드 유래 DNA probe에 의한 Erwinia carotovora subsp. carotovora 특이적 검출)

  • Kang, Hee-Wan;Go, Seung-Joo;Kwon, Soon-Wo
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.164-170
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    • 1998
  • This study was carried out to develop DNA probe for specific detection of Erwinia carotovora subsp. carotovora. Universal rice primer (URP, 20 mer) developed from repetitive sequence of rice was applied for producing PCR DNA fingerprints of Erwinis spp. In E. carotovora subsp. carotovora strains, primer URP2F amplyfied polymorphic bands which are distinguisable from other Erwinia spp. A PCR band of 0.6 kb selected from PCr polymorphic bands of E. carotovora subsp. carotovora strains was cloned and evaluated as a diagnostic DNA probe. Among 28 bacterial strains including 22 Erwinia spp, the probe (pECC2F) only hybridized to total DNAs from e. carotovora subsp. carotovora strains and E. carotovora subsp. wasabiae, but sizes of hybridized bands were different between these subspecies, 10.0 kb and 3.5 kb respectively. In dot blot assays using probe pECC2F, as few as 103 colony forming units (CFU) of E. carotovora subsp. carotovora could be detected in a suspension containing about 1$\times$103 CFU of soil bacteria.

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Cleaved Amplified Polymorphic Sequence and Amplified Fragment Length Polymorphism Markers Linked to the Fertility Restorer Gene in Chili Pepper (Capsicum annuum L.)

  • Kim, Dong Sun;Kim, Dong Hwan;Yoo, Jae Hyoung;Kim, Byung-Dong
    • Molecules and Cells
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    • v.21 no.1
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    • pp.135-140
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    • 2006
  • Cytoplasmic male sterility (CMS) in plants, which is due to failure to produce functional pollen, is a maternally inherited trait. Specific nuclear genes that suppress CMS, termed fertility restorer (Rf) genes, have been identified in several plants. In this study, Rfl-inked molecular markers in pepper (Capsicum annuum L.) were detected by bulked segregant analysis of eight amplified fragment length polymorphisms (AFLPs). Only AFRF8 was successfully converted to a cleaved amplified polymorphic sequence (CAPS) marker. This was named AFRF8CAPS and genotype determination using it agreed with that obtained with the original AFRF8. A linkage map with a total size of 54.1 cM was constructed with AFRF8CAPS and the seven AFLP markers using the Kosambi function. The AFRF8CAPS marker was shown to be closest to Rf with a genetic distance of 1.8 cM. These markers will be useful for fast and reliable detection of restorer lines during $F_1$ hybrid seed production and breeding programs in pepper.