• Title/Summary/Keyword: pKU10

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Characteristics of the R plasmid pKU10 isolated from Pseudomonas putida (Pseudomonas putida에서 분리한 플라스미드 pKU 10의 특성)

  • 임영복;이영록
    • Korean Journal of Microbiology
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    • v.25 no.4
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    • pp.282-289
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    • 1987
  • The characteristics of the plasmid pKU10 isolated from Pseudomonas putida KU816 were investigated and its restriction map was constructed. The pKU10 plasmid was a small R plasmid carrying genes for resistance to ampicillin, tetracyclin, and chloramphenicol, and cured by treatment with mitomycin C. The molecular size of pKU10 was estimated to be 9.4Kb. Pseudomonas strains and E. coli cells could be transformed for antibiotic resistance characters specified by pKU10 plasmid DNA. By incompatibility test with other plasmids, pKU10 is grouped into IncP-1. EcoRI, XhoI, SalI, BglII, and SmaI cleaved pKU10 once, while PstI cleaved at two sites, and HindIII cleaved at six sites. The restriction map was constructed by partial and complete digestion of the purified plasmid DNA with single, double, or triple restriction enzymes. Thus, pKU10 is expected to be used for a cloning vector in Pseudomonas cells.

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Characterization of SAL plasmid isolated from Pseudomonas putida (Pseudomonas putida에서 분리한 SAL 플라스미드의 특성)

  • 김희윤;임영복;이영록
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.9-16
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    • 1987
  • Three strains of bacteria utilizing salicylate, KU801(pKU5, pKU8), KU803(pKU6, pKU9), and KU806(pKU7, pKU10), were selected from the isolates and identified as Pseudomonas putida. By agarose gel electrophoresis, it was found that the strains had two plasmids each. All three strains were resistant to antibiotics such as ampicillin, tetracyclin, and chloramphenicol, and did not utilize other aromatic and aliphatic hydrocarbons examined except salicylate. The plasmids (pKU5, pKU6, and pKU7) of larger molecular weight were cured by treatment with mitomycin C and frequencies of curing were 0.4%, 1.67%, and 0.75%, respectively. Cured strains did not degrade salicylate and still had antibiotic resistances, which were identical with wild strains. The genes for salicylate degradation were proved to be enclded on thier plasmids. The molecular weights of pKU5 and pKU6 were estimated as 103.5Md, and that of pKU 7 as 101 Md. The new SAL plasmids, pKU5, pKU6, and pKU7 were transferred to P. putida and P. aeruginosa, but not to E. coli.

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Restriction Map of the R Plasmid pKU10 in Pseudomonas putida (Pseudomonas putida에 내재하는 Plasmid pKU10의 제한지도)

  • 전성희;임영복;심웅섭;이영록
    • Korean Journal of Microbiology
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    • v.29 no.4
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    • pp.226-229
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    • 1991
  • In our laboratory a R plasmid pKU10 was isolated from Pseudomonas and its characteristics were investigated. In this study, as a basic work to improve its utility as a cloning vehicle, restriction patterns of pKU10 were analyzed for other various restriction enzymes in addition to restriction evdonucleases previously examined. As a result, pKU10 DNA has two cleavage sites for ClaI and HpaI, and three sites for AvaI. The restriction map of pKU10 was supplemented with AvaI, ClaI, and HpaI. From the result of this experiment, the usefulness of PKU10 as a cloning vector in Pseudomonas will be enhanced by constructions of mini-plasmid or hybrid plasmids.

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Morphological and Molecular Identification of Penicillium islandicum Isolate KU101 from Stored Rice

  • Oh, Ji-Yeon;Kim, Eui-Nam;Ryoo, Mun-Il;Kim, Ki-Deok
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.469-473
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    • 2008
  • We have previously obtained a representative isolate KU101 of the predominant Penicillium species from rice under indoor storage conditions. In this study we attempted to characterize isolate KU101 using its morphological and molecular characteristics. When the micro- and macroscopic characteristics of isolate KU101 were compared with the P. islandicum reference isolate KCCM 34763, isolate KU101 was generally identical to those of isolate KCCM 34763, however, isolate KU101 grew faster and produced more orange to red pigments than isolate KCCM 34763. In a molecular-based identification, the nuclear sequence of the ITS1-5.8S-ITS2 region of isolate KU101 was most closely related to that of P. islandicum. Therefore, these results indicated that isolate KU101 from stored rice could be identified as P. islandicum, some isolates of which are known to produce mycotoxins.

A genetic map of the R-factor pKU10 isolated from pseudomonas putida (Pseudomonas putida에서 분리한 R-factor pKU 10의 유전자 지도)

  • 임영복;민경미;이영록
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.167-172
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    • 1988
  • A genetic map of the IncP-1 group plasmid pKU10 has been prepared through the construction of recombinant plasmids containing various fragments of pKU10. Phenotypic analysis of these derivatives has identified the location of genes encoding resistance to ampicillin, tetracyclin, and chloramphenicol. The region involved in conferring resistance to ampicillin was located around two PstI sites that are 1.0Kb apart. The tetracyclin resistance gene was mapped on the region of HindIII E fragment and a part of HindIII D fragment, and the determinant for chloramphenicol resistance gene was localized on HindIII D fragment.

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Pseudomonas sp. 의 균주개발에 유용한 클로닝 백터 pKU11 의 조립

  • 강형일;고상근;이영록
    • Korean Journal of Microbiology
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    • v.30 no.5
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    • pp.410-414
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    • 1992
  • Numerical identification was carried out for an isolate of Streptomyces strain producing the extracellular p-lactamase inhibitor. Fifty taxonomic unit characters were tested and the data were analyzed numerically using the TAXON program. The isolate was identified to the major cluster 5 of Streptomyces and it was best matched to Streptomyces omiyaensis which is a synonym of Streptomyces exfoliatus. Therefore, it was concluded that the isolate was identified to be a strain (SMF 19) of Streptomyces exjbliatus.

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Isolation and Identification of Lactic Acid Bacteria from Spent Mushroom Substrate for Silage Making and Determination of Optimal Medium Conditions for Growth

  • Kim, Young-Il;Kwak, Wan-Sup
    • Journal of Animal Science and Technology
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    • v.54 no.6
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    • pp.435-442
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    • 2012
  • This study was conducted to isolate and identify the lactic acid bacteria (LAB) from spent mushroom substrates (SMS) for the effective anaerobic fermentation to utilize SMS as an animal feed and to determine the optimal medium conditions for their growth. At first, a total of 23 strains were isolated from the ensiled SMS based on the LAB counts and pH tested. Then, a total of 16 strains which rapidly produce lactate and decreased the pH, were selected for a screening test. The optical density (OD), pH, and yellow clear zone were tested for the selected 16 strains. Among the strains, KU5 strain had wider yellow clear zone and lower pH and KU13 strain had higher OD at 24 hr of incubation and wider yellow clear zone compared to other strains and control strain (Lactobacillus plantarum KCCM 12116). Accordingly, KU5 and KU13 strains were finally selected. The KU5 and KU13 were identified as Lactobacillus plantarum by the 16S rRNA sequencing. The KU5 strain was named as Lactobacillus plantarum KU5, and the KU13 strain was named as Lactobacillus plantarum KU13. Lactobacillus plantarum KU5 and Lactobacillus plantarum KU13 were registered at the National Center for Biotechnology Information (NCBI). Access number of Lactobacillus plantarum KU5 was HQ542227 and that of Lactobacillus plantarum KU13 was HQ542228. The optimal medium conditions for growth of KU5 and KU13 were soybean meal 2% and formulated feed 2%, respectively.

Isolation and characterization of corynebacteria-E. coli shuttle vector pKU6 from coryneform bacteria (Corynebacteria-E. coli shuttle vector pKU6의 분리 및 확인)

  • 허태린;이진우;이세영
    • Korean Journal of Microbiology
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    • v.22 no.4
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    • pp.249-255
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    • 1984
  • To develop the host-vector system for industrial Coryneform bacteria that seemed to be the most suitable microorganisms for molecular breeding of genes involved in the production of amion acids, nucleotides, and other products of industrial interest, broad host range E. coli plasmid R 1162 DNA was transformed into Brevibacterium ammoniagenes and the plasmids pKU6 isolated from a transformant was physically characterized. All other plasmids from the transformed cells except pKU6 exsisted as multimeric forms in Brevibacterium ammoniagenes. The plasmid DNA was retransformed into Corynebacterium glutamicum with a high frequency ($1.32{\times}10^{-1}$ per cell) and maintained stably both in Brevibacterium ammoniagenes and Corynebacterium glutamicum after 100 generations of cultures with 25-30 copy number per cell. The size of both plasmid pKU6 and plasmid R1162 were the same and restriction maps by EcoR I, Ava I, Pst I, Pvu II and Hinc II were also similar.

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First Detection of Penicillium fellutanum from Stored Rice in Korea

  • Oh, Ji-Yeon;Sang, Mee-Kyung;Lee, Ho-Joung;Ryoo, Mun-Il;Kim, Ki-Deok
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.216-221
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    • 2011
  • A representative isolate KU53 of the predominant Penicillium species was obtained from rice samples from rice processing complexes of National Agricultural Cooperative Federation in Korea. In this study, isolate KU53 was identified by its morphological and molecular characteristics. The macro- and microscopic characteristics of isolate KU53 were compared with the P. fellutanum reference isolate KCTC16913 on different media; isolate KU53 was generally identical to those of the reference isolate KCTC16913. In a molecular-based identification, the ${\beta}$-tubulin and translation elongation factor 1-alpha sequences of isolate KU53 was most closely related to those of P. fellutanum. Thus, isolate KU53 from stored rice could be identified as P. fellutanum, some isolates of which are known to produce mycotoxin-related metabolites. To our knowledge, this is the first detection of P. fellutanum from stored rice in Korea.

Morphological Characteristic Regulation of Ligninolytic Enzyme Produced by Trametes polyzona

  • Lueangjaroenkit, Piyangkun;Teerapatsakul, Churapa;Chitradon, Lerluck
    • Mycobiology
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    • v.46 no.4
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    • pp.396-406
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    • 2018
  • A newly isolated white rot fungal strain KU-RNW027 was identified as Trametes polyzona, based on an analysis of its morphological characteristics and phylogenetic data. Aeration and fungal morphology were important factors which drove strain KU-RNW027 to secrete two different ligninolytic enzymes as manganese peroxidase (MnP) and laccase. Highest activities of MnP and laccase were obtained in a continuous shaking culture at 8 and 47 times higher, respectively, than under static conditions. Strain KU-RNW027 existed as pellets and free form mycelial clumps in submerged cultivation with the pellet form producing more enzymes. Fungal biomass increased with increasing amounts of pellet inoculum while pellet diameter decreased. Strain KU-RNW027 formed terminal chlamydospore-like structures in cultures inoculated with 0.05 g/L as optimal pellet inoculum which resulted in highest enzyme production. Enzyme production efficiency of T. polyzona KU-RNW027 depended on fungal pellet morphology as size, porosity, and formation of chlamydospore-like structures.