• Title/Summary/Keyword: restriction

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Complementary DNA Cloning and Restriction Mapping of Nuclear Inclusion Body and Coat Protein Genes of Turnip Mosaic Virus-Ca Strain Genomic RNA (순무모자이크 바이러스 Ca계통 핵봉입체와 외피단백질 유전자의 cDNA 클로닝 및 제한효소 지도작성)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.235-239
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    • 1994
  • Viral RNA was extracted from purified Chinese cabbage strain of turnip mosaic virus (TuMV-Ca) from infected leaves of turnip. Polyadenylated genomic viral RNA was recovered by oligo (dT) cellulose column chromatography and used as a template for the synthesis of complementary DNA (cDNA). Recombinant plasmids contained cDNA ranged from about 900 bp to 2, 450 bp were synthesized. Among the selected 41 transformants, pTUCA31 and pTUCA35 had over 2 Kbp cDNA insert. Restriction endonuclease patterns of the clones examined were very similar among them. Clones pTUCA23 and pTUCA31 were overlapped with pTUA35. The longest clone pTUCA35, encoding 3'-end, showed that it contained two sites for EcoRI, and one site for BamHI, ClaI, HincII, SacI and XbaI, respectively. The restriction mapping indicated that the clone pTUCA35 contained partial nuclear inclusion body gene, complete coding region of the coat protein and 3' untranslated region of TuMV-Ca genomic RNA.

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Monitoring of Microorganisms Added into Oil-Contaminated Microenvironments by Terminal-Restriction Fragment Length Polymorphism Analysis

  • JUNG SEONG-YOUNG;LEE JUNG-HYUN;CHAI YOUNG-GYU;KIM SANG-JIN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1170-1177
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    • 2005
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to monitor inoculated oil-degrading microorganisms during bioremedial treatability tests. A pair of universal primers, fluorescently labeled 521F and 1392R, was employed to amplify small subunit rDNA in order to simultaneously detect two bacterial strains, Corynebacterium sp. IC10 and Sphingomonas sp. KH3-2, and a yeast strain, Yarrowia lipolytica 180. Digestion of the 5'-end fluorescence/labeled PCR products with HhaI produced specific terminal-restriction fragments (T-RFs) of 185 and 442 bases, corresponding to Corynebacterium sp. IC10 and Y. lipolytica 180, respectively. The enzyme NruI produced a specific T-RF of 338 bases for Sphingomonas sp. KH3-2. The detection limit for oildegrading microorganisms that were inoculated into natural environments was determined to be $0.01\%$ of the total microbial count, regardless of the background environment. When three oil-degrading microorganisms were released into oil-contaminated sand microenvironments, strains IC10 and 180 survived for 35 days after inoculation, whereas strain KH3-2 was detected at 8 days, but not at 35 days. This result implies that T-RFLP could be a useful tool for monitoring the survival and relative abundance of specific microbial strains inoculated into contaminated environments.

Genetic Relationships of Cattle Breeds Assessed by PCR-RFLP of the Bovine Mitochondrial DNA D-loop Region

  • Yoon, Du Hak;Lee, Hak Kyo;Oh, Sung Jung;Hong, Ki Chang;Jeon, Gwang Joo;Kong, Hong Sik;Lee, Jun Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1368-1374
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    • 2005
  • To investigate the genetic relationships among various cattle breeds, bovine mtDNA D-loop region was used in 411 animals of 18 cattle breeds, including 8 Asian Bos taurus, 7 European Bos taurus, 1 Asian Bos indicus, and 2 African Bos indicus. The size of amplified PCR products from mtDNA D-loop region was 964 bp and the products were digested by 15 different restriction enzymes. Two different band patterns were identified in eight restriction enzymes (BstXI, Hae III, Msp I, Apa I, Taq I, Alu I, BamH I, EcoN I) and the rest of restriction enzymes showed more than 3 different band patterns among which Apo I and MspR9 resulted in 7 different restriction patterns. The genotypes, number of haplotype, effective number of haplotype, and degree of heterozygosity were analyzed. Based on all the PCR-RFLP data, different haplotypes were constructed and analyzed for calculating genetic distances between these breeds using Nei's unbiased method and constructing a phylogenetic tree.

Survey of Bedrock Quality and Suitable Site for Crushed Stones using GIS in the Wondeung Area (GIS를 이용한 원등일대의 기반암품질과 골재개발적지 분석)

  • Lee, Jin-Young;Yang, Dong-Yun;Kim, Ju-Yong;Hong, Sei-Sun;Yun, Hyun-Soo
    • Journal of Korean Society for Geospatial Information Science
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    • v.13 no.4 s.34
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    • pp.47-58
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    • 2005
  • Investigation of bedrock quality and the legal restriction for developing the aggregate resources were carried out with GIS in the Wondeung area Gokseong-gun, Jeollanam-do province. Firstly, we surveyed the exposed bedrocks in the field and examined in physical properties in the lab. Secondly we analyzed the legal restriction by using GIS based on bufferzone and viewshed analysis. At the results of this study, the area can be used totally as the crushed stones. However the suitability sites of crushed stone have various regulations. So the legal restriction is the principal standard to develop the crushed stone in the Wondeung area. Suitable site for crushed stones are about 30% in the Wondeung area.

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End-to-end Structural Restriction of α-Synuclein and Its Influence on Amyloid Fibril Formation

  • Hong, Chul-Suk;Park, Jae Hyung;Choe, Young-Jun;Paik, Seung R.
    • Bulletin of the Korean Chemical Society
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    • v.35 no.12
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    • pp.3542-3546
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    • 2014
  • Relationship between molecular freedom of amyloidogenic protein and its self-assembly into amyloid fibrils has been evaluated with ${\alpha}$-synuclein, an intrinsically unfolded protein related to Parkinson's disease, by restricting its structural plasticity through an end-to-end disulfide bond formation between two newly introduced cysteine residues on the N- and C-termini. Although the resulting circular form of ${\alpha}$-synuclein exhibited an impaired fibrillation propensity, the restriction did not completely block the protein's interactive core since co-incubation with wild-type ${\alpha}$-synuclein dramatically facilitated the fibrillation by producing distinctive forms of amyloid fibrils. The suppressed fibrillation propensity was instantly restored as the structural restriction was unleashed with ${\beta}$-mercaptoethanol. Conformational flexibility of the accreting amyloidogenic protein to pre-existing seeds has been demonstrated to be critical for fibrillar extension process by exerting structural adjustment to a complementary structure for the assembly.

A Molecular Marker Specific to Metarhizium anisopliae var. majus

  • YOON, CHEOL-SIK;GI HO SUNG;JAE MO SUNG;JAEANG OON LEE
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.334-339
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    • 1999
  • More innovative molecular markers were investigated for rapid and consistent differentiation of Metarhizium anisopliae var. majus from M. anisopliae var. anisopliae. A total of 28 isolates were obtained from various countries and hosts: 13 isolates of M. anisopliae var. anisopliae, 12 isolates of M. anisopliae var. majus, and 3 isolates of M. anisopliae collected in Korea. This study involved restriction enzyme digestions of a PCR product amplified from nuclear internally transcribed spacer (ITS) and a portion of the 28S rDNA regions. Among 11 different restriction enzymes used in this study, MboⅠ digestion particularly produced a restriction pattern that had characteristics of M. anisopliae var. majus. This restriction pattern was consistent in all isolates of M. anisopliae var. majus regardless of their geographic origins and insect hosts. Mapping experiments revealed that MboⅠ sites of M. anisopliae var. majus are identical to those of M. anisopliae var. anisopliae with an exception for the presence of an additional site in the PCR product. Results from this study provide an additional method for identification and differentiation of isolates of these two varieties of M. anisopliae for use in the field and laboratory experiments.

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Authentication of Salted-dried Fish Species Using Polymerase Chain Reaction-Single Strand Conformational Polymorphism and Restriction Analysis of Mitochondrial DNA

  • Kim, Joo-Shin;Chu, Kin Kan Astley;Kwan, Hoi Shan;Chung, Hau Yin
    • Fisheries and Aquatic Sciences
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    • v.11 no.3
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    • pp.133-139
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    • 2008
  • Molecular techniques, including restriction fragment length polymorphism(RFLP) and polymerase chain reaction-single strand conformational polymorph isms(PCR-SSCP), were developed to identify salted, dried threadfin(Eleutheronema tetradactylum) and white herring(Ilisha elongata) fish. Using PCR with universal primers, conserved 367-bp fragments of the cytochrome b gene were amplified from fresh fish samples and sequenced. The sequences were then searched for specific restriction sites. The digestion of the PCR products with the endonucleases AvaI, FokI, MboII, and MspI generated RFLP, which was used to identify the commercial products. Similarly, the amplified PCR-SSCP products were developed and the products tested. Overall, similar patterns were found in the majority of the fresh and processed products. Based on the results, both RFLP and PCR-SSCP were useful in determining and validating the authenticity of the fish species used to prepare the commercial salted, dried products. A similar approach can be applied to other species.

Purification and Characterzation of a Restriction Endonuclease from Pseudomonas syringae pv.phaselicola (Pseudomonas syringe pv. phaseolicola로 부터 제한효소의 분리정제 및 특성)

  • Bae, Moo;Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.485-490
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    • 1994
  • A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

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Effects of Knee Extension Exercise Using Blood Flow Restriction on the Thickness and Balance Ability of Tendons

  • Park, Jae-Cheol;Kim, Yong-Nam
    • The Journal of Korean Physical Therapy
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    • v.30 no.2
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    • pp.41-46
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    • 2018
  • Purpose: This study examined the effects of knee extension exercise using blood flow restriction on the changes in the thickness and static balance ability of the tendon of the rectus femoris muscle and achilles tendon. Methods: A total of 30 subjects was were divided into two groups of 15 subjects each. The changes in the thickness and balance of their tendons were measured by ultrasonography and balance equipment. The measurements were taken three times: before the experiment, 4 weeks after, and 8 weeks after. The results were analyzed by repeated measure ANOVA, one-way ANOVA was conducted in cases where there were time-to-group interactions in the intra-individual effects test. Results: Significant differences in the thickness of the tendon of the rectus femoris muscle and Achilles tendon were found between the groups, over time, and in the time-to-group interactions (p<0.05). The changes in balance in both feet when the supporting positions between eye-open and eye-closed states were significantly different in the time-to-group interactions (p<0.05), but were not different between the groups (p>0.05). Conclusion: Knee extension exercise using blood flow restriction leads to significant differences in the increase in tendon thickness and changes in balance, indicating that the results of this study can be utilized as basic data for future studies and for rehabilitation treatment at clinics.

Characterization of infectious bursal disease viruses isolated in Korea using RT/PCR and RFLP analysis (RT/PCR과 RFLP 분석에 의한 Infectious bursal disease virus(국내분리주)의 특성 규명)

  • Kwon, Hyuk-moo;Kim, Dae-kyu;Seong, Hwan-woo
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.104-110
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    • 1999
  • Field infectious bursal disease viruses (IBDVs) were isolated from IBDV-suspected commercial chickens. The variable region in VP2 gene of six Korean IBDV isolates (K-IBDVs) and IBD vaccines was examined using the reverse transcriptase / polymerase chain reaction-restriction fragment length polymorphism (RT/PCR-RFLP) assay. With all K-IBDVs and vaccine IBDVs, a 474-bp fragment of the VP2 gene was amplified and tested with various restriction enzymes. Restriction enzymes BstNI and StyI differentiated K-IBDV isolates and IBD vaccines into four groups. Restriction enzyme profiles of K-IBDV isolates were different from them of IBD vaccines. K-IBDV isolates except for 310 isolate had specific SspI and TaqI recognition sites, which were recognized in highly virulent IBDVs, but IBD vaccines had no those sites. This study showed that RT/PCR-RFLP assay was thought to be valuable tool for differentiation of IBDVs and identification of highly virulent IBDV.

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