• Title/Summary/Keyword: restriction analysis

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Restriction Analyses of PCR Amplified Partial SSU Ribosomal DNA to Distinguish Arbuscular Mycorrhizal Fungi from Other Fungi Colonizing Plant Roots

  • Lee, Jae-Koo;Tae, Moon-Sung;Eom, Ahn-Heum;Lee, Sang-Sun
    • Mycobiology
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    • v.31 no.2
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    • pp.68-73
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    • 2003
  • Roots of Glycine max and Miscanthus sinensis and soil samples were collected from various field sites at Goesan, Chungbuk in Korea. Microscopic observations of the roots indicated high colonization rates of both arbuscular mycorrhizal fungi(AMF) and other fungi. The partial small subunit of ribosomal DNA genes were amplified with the genomic DNA extracted from their roots by nested polymerase chain reaction(PCR) with universal primer NS1 and fungal specific primers AML Restriction fragment length polymorphism(RFLP) was analyzed using the combinations of three restriction enzymes, HinfI, AluI and AsuC21. Nucleotides sequence analysis revealed that ten sequences from Miscanthus sinensis and one sequence from Glycine max were close to those of arbuscular mycorrhizal fungi. Also, 33% of total clones amplified with NS31-AM1 primers from M. sinensis and 97% from G. max were close to Fusarium oxysporum or other pathogenic fungi, and they were successfully distinguished from AME Results suggested that these techniques could help to distinguish arbuscular mycorrhizal fungi from root pathogenic fungi in the plant roots. Especially, DNA amplified by these primers showed distinct polymorphisms between AMF and plant pathogenic species of Fusarium when digested with AsuC21.

Polymorphisms in Exon 2 of MHC Class II DRB3 Gene of 10 Domestic Goats in Southwest China

  • Zhao, Yongju;Xu, Huizhong;Shi, Lixiang;Zhang, Jiahua
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.6
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    • pp.752-756
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    • 2011
  • Polymorphism of the second exon of the caprine leukocyte antigen-DRB3 gene (CLA-$DRB3^*02$) was investigated in this study. The 285 bp PCR product of 258 individuals from 10 domestic goat breeds in Southwest China was digested with restriction endonucleases PstI and HaeIII and then genotyped. Three alleles and 4 restriction digestion profiles were distinguished by digestion of the PCR fragment by PstI, and 8 alleles and 13 genotypes by HaeIII. For HaeIII restriction enzyme sites, the Chi-square ($X^2$) test showed that all goat breeds in this study did not fit with the Hardy-Weinberg equilibrium (p<0.01 or p<0.05). The highly polymorphic nature of CLA-$DRB3^*02$ was demonstrated and the ranges of gene heterozygosity (He) and polymorphism information content (PIC) were 0.36-0.63 and 0.32-0.55, respectively. Clustering analysis showed that the 10 goat breeds clustered into two groups and Dazu Black goat had a close genetic relationship with Chengdu Grey, Jintang Black and Nanjiang Yellow goats.

The Change of Electrolytes Composition in Skeletal Muscles by Protein Restriction and Rehabilitation (단백질의 제한과 회복으로 인한 골격 근육내의 성분변화)

  • Kim, Jee-Hyun;Kim, Sook-He
    • Journal of Nutrition and Health
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    • v.14 no.4
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    • pp.151-161
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    • 1981
  • The objective of this study was to compare the electrolytes contents in skeletal muscles of two different groups of rats, one for control fed 20% casein diet, and the other experimental group recovered from 7% casein diet as protein restriction feeding for two weeks at the beginning of this experiment. Two different comparison criteria were used in this study, one for comparison between animals at the same age groups and the other between groups weighing same body weight as the control groups. Food consumption of animals for experimental and control groups were ad libitum. Following measurement and samples were collected: body weight, five different muscles-anterior tibialis (A.T.) extensor digitorium longus (E.D.L.) soleus, plantaris, gastrocnemius-for electrolytes and protein analysis. The experimental groups showed lower body weight than that of control group. Same trend was showen in protein content in five different muscles. Magnesium and potassium content in four muscles except gastrocnemius showed lower and sodium higher in experimental groups than those in control group at fourteenth day, after recovery from protein restriction, electrolytes content change showed differently in four muscles. Magnesium and potassium contents were increased as protein content in diet. Sodium content decreased as one week intake of recovery diet started, thereafter it was rather slowly increased. Phosphorus content in gastrocnemius at the second week in experimental group was slightly lower than that of control group, and it recovered rapider while at the same body weight group it was higher in experimental group.

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Molecular cloning and restriction endonuclease mapping of homoserine dehydrogenase gene (HOM6) in yeast saccharomyces cerevisiae (Aspartate계 아미노산 대사 효모 유전자 HOM6의 cloning 및 구조분석)

  • 김응기;이호주
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.357-363
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    • 1986
  • Synthesis of threonine and methionine in yeast, Saccharomyces cerevisiae shares a common pathway from aspartate via homoserine. HOM6 gene encodes homoserine dehydrogenase (HSDH) which catalyzes the inter-conversion of beta-aspartate semialdehyde and homoserine. The level of HSDH is under methionine specific control. A recombinant plasmid (pEK1: 13.3kb), containing HOM6 gene, has been isolated and cloned into E. coli by complenemtary transformation of a homoserine auxotrophic yeast strain M-20-20D (hom6, trp1, ura3) to a prototrophic M20-20D/pEK1, using a library of yeast genomic DNA fragments in a yeast centromeric plasmid, YCp50(8.0kb). Isolation of HOM6has been primarily confirmed by retransformation of the original yeast strain M20-20D, using the recombinant plasmid DNA which was extracted from M20-20D/pEK1 and subsequently amplified in E. coli. Eleven cleavage sites in the insery (5.3kb) have been localized through fragment analysis for 8 restriction endonucleases; Bgl II(2 site), Bgl II(1), Cla I(3), Eco RI(1), Hind III(2), Kpn I (1), Pvu II(1) and Xho I(1).

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INTRAFAMILIAL TRANSMISSION OF ACTINOBACILLUS ACTINOMYCETEMCOMITANS IN KOREAN POPULATIONS (한국인에 있어서 Actinobacillus actinomycetemcomitans의 가족내 전이양상에 관한 연구)

  • Jeong, Young-In;Kim, Sung-Jo;Choi, Jeom-Il
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.679-693
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    • 1995
  • The present study has been performed to see the intrafamilial transmission of periodontopathic organism Actinobacillus actinomycetemcomitans in Koreans having various froms of periodontal diseases. 17 clinical isolates from 8 periodontal patients and 20m clinical isolates from their 8 family members were grown anaerobically for the serotyping and the extraction of genomic DNA. The DNA was digested with restriction endonucleases (EcoRI+HindIII) and plasmid pAA 2097(kindly provided by Dr. DiRienzo, Univ. of Pennsylvania) including 4.7kb-size randlomly clone probe for restriction length pleomorphism analysis(RFLP). RFLP patterns of reference serotypes a, b, c, d, and e were used as the genotypes A, B, C, D, and E, respectively for comparison of genotypes of clinical isolates. 28 out of total 37 clinical isoltes belonged to either one of 5 basic gentotypes and 9 remaining isolats did not fall into any types, and hence were designate as non-type(NT). Genotype C were the most frequently found one(35.1%) and genotype B has not isolated. Intrafamilial transmission of bacteria between spouses, brothers and sisters, and parents and their offsprings, resepctively could well be demonstrated by comparing RFLP patterns. There were not any specific genotypes which showed predominance over others in terms of transmission.

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Isolation, Restriction Mapping, and Promoter Sequence Analysis of an Isoperoxidase Gene from Korean-Radish, Raphanus sativus L.

  • Park, Jong-Hoon;Kim, Soung-Soo
    • BMB Reports
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    • v.29 no.1
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    • pp.52-57
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    • 1996
  • A specific DNA fragment from Korean radish (Raphanus sativus L.) was amplified by performing PCR with oligonucleotide primers which correspond to the highly conserved regions of plant peroxidases. The size of the PCR product was ca. 400 bp, as expected from the known plant peroxidase genes. Comparison of the nucleotide and deduced amino acid sequences of the PCR product to those of other plant peroxidase-encoding genes revealed that the amplified fragment corresponded to the highly conserved region I and III of plant peroxidases. By screening a genomic library of Korean radish using the amplified fragment as a probe, two positive clones, named prxK1 and prxK2, were isolated. Restriction mapping studies indicated that the 5.2 kb Sail fragment of the prxK1 clone and the 4.0 kb EcoRI fragment of the prxK2 clone encode separate isoperoxidase genes. Analyses of the promoter region of the prxK1 clone shows that putative CAAT box, CMT box, and TGA1b binding sequence (5' TGACGT) are present 718 bp upstream from the start codon.

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Some Theoretical Foundations on the Necessities and Functions of Global Electronic Transactions Act (전자무역 활성화를 위한 글로벌 전자무역거래법의 요건과 역할기능의 이론적 기초)

  • Kim, Ki-Sun
    • THE INTERNATIONAL COMMERCE & LAW REVIEW
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    • v.17
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    • pp.129-146
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    • 2002
  • The electronic technology development have occurred in the face of existing legal barriers to legal efficacy of computer information goods, and the liberating promise of electronic transactions cannot fully realized unless there is predictability in the legal rules that govern such transactions. This study analyzes some theoretical fundamentals of the Act. First, it proposes that the Act clarify and set forth uniform legal principles applicable to computer information transactions. Secondly, it suggests that if the individual is risk averse, the acceptance set for electronic transactions will be a convex set, and that the application of the Act will make the acceptance set more expanded by lowering the probability of conflicts and by downsizing the risk averness. Thirdly, it also suggest that through the mothod of contingent commodities analysis, the application of the Act by means of its restricted regulations will give more expected utility than the absence of the Act. Fourthly, it derives some implications that the degree of legitimate restriction will be affected by the objective risk inherent to the electronic transactions, and the individual's subjective risk-averseness. Finally, it concludes that harmonization of restriction and protection of individual's rights in electronic transaction process will be a necessary condition for more efficient body of law from the law-economic perspectives.

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Characterization of Plasmid Encoding a Heat-labile Enterotoxin Originated from an Human Escherichia coli Strain O15: H11. (사람에서 유래한 장독성 대장균의 이열성장독소와 관련된 Plasmid 특성)

  • Rhyu, Mun-Gan;Kim, Gum-Ryong;Chang, Woo-Hyun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.47-52
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    • 1986
  • A heat-labile enterotoxin and no heat-stable enterotoxin producing($LT^+ST^-$) plasmid (110 kilobases in size) was isolated from an enterotoxigenic Escherichia coli of human strain O15:H11 and used for analysis of the $LT^+$ deoxyrionucleic acid region using recombinant DNA technology. A DNA segment containing the $LT^+$ DNA region which was one restriction endonuclease BamHl fragment(6.2 kb in size) was joind to a small multicopy plasmid, pUC9. E. coli K-12 strain, JM103 harboring the chimeric plasmid produced greater amounts of LT than did the enterotoxigenic E. coli O15:H11 strain. The BamHl fragment was further digested with various restriction endonucleases and contained no HindIll restriction site which is an essential in $LT^+ST^+$ plasmid. The detailed DNA sequencing of this $LT^+ST^-$ plasmid is required.

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Genetic Diversity of Korean Cyanobacteria determined by DNA polymorphisms within the Phycocyanin Locus (Phycocyanin locus내의 DNA Polymorphism에 의한 한국산 Cyanobacteria의 유전적 다양성)

  • 박진숙;권주리;유순애
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.249-253
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    • 2000
  • The genetic diversity among Korean cyanobacteria was assessed by restriction fragment length polymorphism(RFLP) analysis of PCR products from the phycocyanin locus. Strains of all the genera tested were successfully amplified, and the size of amplified fragments was approximately 700bp. The restriction patterns generated by AluI, MspI, and HaeIII were conserved for strains within each of genera studied and were specific to the genus level. Intrageneric delineation of strains was revealed by the enzyme, CfoI for members of genera Anabeana and Synechocystis. Phenogram derived from the different RFLP patterns revealed a coherent cluster among Anabeana, Chlorogloea, and Synechocystis strains. PC-RFLP methods provided useful tools for classification of cyanobacteria.

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Identification of Aster Yellows Phytoplasma in Dendranthema grandiflorum

  • Chung, Bong-Nam;Park, Gug-Seoun;Kim, Hyun-Ran;Park, Yong-Mun
    • The Plant Pathology Journal
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    • v.17 no.1
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    • pp.57-61
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    • 2001
  • Phytoplasmas were identified from two chrysanthemum (Dendranthema grandiflorum) plants showing different symptoms ; one with stusting, rosette, and excessive branching (Ph-ch1), and the other with stunting and chlorosis (Ph-ch2). Electron microscopy of midrib of the plants with the symptoms revealed that numerous phytoplasmas were localized in the phloem cells. The disease was transmitted from infected plants to healthy ones by grafting. Phytoplasma-specific DNA was detected in polymerase chain reaction (PCR) analysis with template DNA extracted from the leaves of Ph-ch1 and Ph-ch2, both of which yielded a same DNA band corresponding to 1.5 kb. Using a specific primer pair (R16F1/R1) synthesized based on aster yellows (AY) phytoplasma, a DNA fragment of 1.1 kb was amplified by PCR. Endonuclease restriction patterns of the 1.1 kb PCR products from Ph-ch1 and Ph-ch2, which were dgeste with each of the restriction endonucleases Sau3A, Hha, Alu and Rsa, were same as those of AY phytoplasma from periwinkle. This suggests that the chrysanthemum plants (Ph-ch1 and Ph-ch2) be infected with a phytoplasma belonging to AY phytoplasma.

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