• Title/Summary/Keyword: plasmid content

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Production System for Biodegradable Polyester Polyhydroxybutyrate by Corynebacterium glutamicum

  • Jo, Sung-Jin;Ooi, Toshihiko;Taguchi, Seiichi
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.352-352
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    • 2006
  • Corynebacterium glutamicum, which is well known as an amino acid fermentation bacterium, has been used as a producer of poly(3-hydroxybutyrate) [P(3HB)]. P(3HB) was synthesized in recombinant C. glutamicum harboring the expression plasmid vector with a strong promoter for cell surface protein gene derived from C. glutamicum and P(3HB) biosynthetic gene operon derived from Ralstonia eutropha. The expression of P(3HB) synthase gene was detected by enzyme activity assay. Intracellular P(3HB) was microscopically observed as inclusion granules and its content was calculated to be 22.5 % (w/w) with molecular weight of $2.1{\times}10^{5}$ and polydispersity of 1.63.

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Genome analysis of Limosilactobacillus fermentum JN2019 applied to tumeric fermentation for animal feed

  • Yoo, Heeseop;Yong, Cheng Chung;Oh, Sejong
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1204-1206
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    • 2021
  • Limosilactobacillus fermentum JN2019, formerly named Lactobacillus fermentum JN2019, was isolated from kimchi. Its genome was completely sequenced using the PacBio RSII sequencing system to explore beneficial phenotypes. In a previous study, L. fermentum JN2019 was used to ferment the by-product of tumeric for use in livestock feed. The 2.3 Mb genome had a high guanine (G) + cytosine (C) content of 50.6% and a 30 kb plasmid. The data will inform the comprehensive understanding of JN2019 and provide insights for potential applications.

Complete genome sequence of a methicillin-resistant Staphylococcus schleiferi strain from canine otitis externa in Korea

  • Lee, Gi Yong;Yang, Soo-Jin
    • Journal of Veterinary Science
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    • v.21 no.1
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    • pp.11.1-11.7
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    • 2020
  • The increase in canine skin and soft tissue infections, such as pyoderma and otitis, caused by Staphylococcus schleiferi strains, is of significant zoonotic concern. In this study, we report the first complete genome sequence for a methicillin-resistant clinical isolate of S. schleiferi (MRSS) designated as SS4, obtained from a dog with otitis externa, in Korea. The genome of SS4 strain was of 2,539,409 bp and presented high G+C content ratio (35.90%) with no plasmid. Comparative analysis of SS4 genome revealed that it is closely related to 2142-05 and 5909-02 strains isolated from the canine skin infections in the USA.

Complete genome and two plasmids sequences of Lactiplantibacillus plantarum L55 for probiotic potentials

  • Bogun Kim;Kiyeop Kim;Xiaoyue Xu;Hyunju Lee;Duleepa Pathiraja;Dong-June Park;In-Geol Choi;Sejong Oh
    • Journal of Animal Science and Technology
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    • v.65 no.6
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    • pp.1341-1344
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    • 2023
  • In this study, we report the complete genome sequence of Lactiplantibacillus plantarum L55, a probiotic strain of lactic acid bacteria isolated from kimchi. The genome consists of one circular chromosome (2,077,416 base pair [bp]) with a guanine cytosine (GC) content of 44.5%, and two circular plasmid sequences (54,267 and 19,592 bp, respectively). We also conducted a comprehensive analysis of the genome, which identified the presence of functional genes, genomic islands, and antibiotic-resistance genes. The genome sequence data presented in this study provide insights into the genetic basis of L. plantarum L55, which could be beneficial for the future development of probiotic applications.

Autonomously Mitochondrial Replicating Sequence of Aspergillus nidulans (Aspergillus nidulans mtDNA의 자가복제절편)

  • 장승환;한동민;장광엽
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.218-225
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    • 1999
  • We isolated the ANRI fragment from Aspergillus nidulons that could autononlously replicate and enhance transformation efficiency about $10^4$ fold compared lo the integrative vector in Saccha,omgcer cerevisioe. In A. nidulans recombinant plasmid pLJ16-4.5 which carries the 4.5 kb EcoRI fragment of ANRI showed a 170-[old increase of transformation efficiency compared to the integrative vector pLJ16 and could be recovered from iransfonnants as an intact form. Estimated copy number of transforming plasmid pLJ16-4.5 was scored as 2 to 3 copies in transformed A. nidulans. Recoinbinant plasinid pILJ16-4.5 is inilotically unstable; being lost Irom 65% of aswual progeny of transformants on selective medium and 90% on complete medium. Southern analysis of transformant DNA showed that the pILJ16-4.5 is maintained in free form. The sequencing data showed that ANRl fragment was originated from mitochondiral DNA of A. nid~ilans and contained high AT content as much as 74.7%. One ARS consensus sequence (A/T)TTr4T(A/G)TTT(AiT). I I ARS-like sequence (agreement 10 of 11) and ABFl binding core consensus sequence (TCN7ACG). Also six gyrase binding core consensus sequence (YRTGNYNNY: y=C or T, R=A or G, N=A, G, C or T) of $\Phi$X174 and SV40 DNA and one b site (CACTTTACC) combining with gyrase in ColEl are shown. ANRl can be developed as a repl&ng plasinid for lransfoimation system in A. nirlulmis.

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Characterization of a Small Cryptic Plasmid from Pseudomonas nitroreducens Strain TX1 (Pseudomonas nitroreducens TX1에 존재하는 작은 플라스미드의 특성 규명)

  • Nguyen, Ngoc Tuan;Lee, Kyoung;Kang, Ju Beom;Huang, Shir-Ly
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.210-215
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    • 2014
  • Pseudomonas nitroreducens TX1 was isolated from a rice field drainage in Taiwan. The bacterium is of special interest because of its capability to use a group of nonionic surfactants such as alkylphenol polyethoxylates even at high concentrations as a sole carbon source. In this study, a small cryptic circular plasmid, pTX1, was characterized from P. nitroreducens TX1. It is 2,286 bp in length with a GC content of 63.3% and harbors three open reading frames, $Rep_{pTX1}$ and functionally unidentified ORF1 and ORF2. The predicted $rep_{pTX1}$ gene product is homologous to Rep proteins of plasmids belonging to the pC194/pUB110 family, which is predominantly found in Gram-positive bacteria and is known to replicate by the rolling-circle mechanism. The copy number of pTX1 was estimated to be about 150 in each cell. Based on the genetic fingerprints and comparison with other plasmids, it is concluded that pTX1 replicates by a rolling circle mechanism which is rarely found for Pseudomonas plasmids.

Thermoresistant properties of bacterioferritin comigratory protein against high temperature stress in Schizosaccharomyces pombe (Schizosaccharomyces pombe에 존재하는 bacterioferritin comigratory protein의 고온 스트레스에 대한 열저항적 성질)

  • Ryu, In Wang;Lee, Su Hee;Lim, Hye-Won;Ahn, Kisup;Park, Kwanghark;Sa, Jae-Hoon;Jeong, Kyung Jin;Lim, Chang-Jin;Kim, Kyunghoon
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.398-405
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    • 2016
  • The Schizosaccharomyces pombe structural gene encoding bacterioferritin comigratory protein (BCP) was previously cloned using the shuttle vector pRS316 to generate the BCP-overexpressing plasmid pBCP10. The present work aimed to evaluate the thermoresistant properties of BCP against high temperature stress using the plasmid pBCP10. When the S. pombe cells were grown to the early exponential phase and shifted from $30^{\circ}C$ to $37^{\circ}C$ or $42^{\circ}C$, the S. pombe cells harboring pBCP10 grew significantly more at both $37^{\circ}C$ and $42^{\circ}C$ than the vector control cells. After 6 h of the shifting to higher incubation temperatures, they contained the lower reactive oxygen species (ROS) and nitrite content, an index of nitric oxide (NO), than the vector control cells. After the temperature shifts, total glutathione (GSH) content and total superoxide dismutase (SOD) activities were much higher in the S. pombe cells harboring pBCP10 than in the corresponding vector control cells. Taken together, the S. pombe BCP plays a thermoresistant role which might be based upon its ability both to down-regulate ROS and NO levels and to up-regulate antioxidant components, such as total GSH and SOD, and subsequently to maintain thermal stability.

Induction of Ginseng Hairy Roots And Their Possible Application To Large Scale Culture

  • Yang, Deok-Chun
    • Plant Resources
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    • v.6 no.1
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    • pp.1-6
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    • 2003
  • Ginseng(Panax ginseng C. A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng hairy roots induced by introducing Ri-plasmid of Agrobacterium rhizogenes into genomic DNA of plant cells show vigorous growth, and the hairy roots produce the same or more saponins than natural ginseng roots. Therefore, hairy roots can be used for commercial purposes. The present study was carried out to induce hairy roots with both active growth and high saponin contents. Numerous hairy roots of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes R1000 A4T in dark condition after one month of culture. About 3 hundred lines of hairy roots were selected according as morphological characters on medium with carbenicillin. After pre-selection of fifteen lines of hairy roots with active growth, KGHR-l and KGHR-8 lines were finally selected which had characters of high content of ginsenoside-Rd and ginsenoside-Re, respectively. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark (22 $^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5L Erlenmeyer flasks, lL roller drums, 10L jar-fermenters, and especially in 20L air-lift culture vessels.

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Transcriptional Regulation of Escherichia coli serC-aroA Operon : Further Support for cAMP-Dependent Expression

  • Sa, Jae-Hoon;Park, Soo-Sun;Lim, Chang-Jin
    • BMB Reports
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    • v.28 no.1
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    • pp.21-26
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    • 1995
  • The Escherichia coli mixed-function serC-aroA operon encodes biosynthethic enzymes for unrelated pathways leading to the syntheses of serine and aromatic amino acids. It has been proposed that the operon is expressed in a cAMP-dependent manner. In this work experiments were performed to investigate the cAMP-dependent expression of the operon. Exogenous cAMP increased ${\beta}$-galactosidase synthesis in the $cya^+$ and cya strains harboring the serC-aroA-lac fusion plasmid. This enhancement was more dramatic in the $cya^-$ strain grown in a minimal medium. In a dot blot assay the serC-aroA mRNA content increased in a concentration-dependent pattern after the addition of exogenous cAMP. The activity of phosphoserine aminotransferase, encoded by the serC gene, apparently increased in E. coli cells after the addition of cAMP. All results obtained confirmed that the expression of the E. coli serC-aroA operon is positively regulated by cAMP at the level of transcription.

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Metabolic Analysis of Poly(3-Hydroxybutyrate) Production by Recombinant Escherichia coli

  • WONG, HENG HO;RICHARD J. VAN WEGEN;JONG-IL CHOI;SANG YUP LEE
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.593-603
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    • 1999
  • Poly(3-hydroxybutyrate) (PHB) production by fermentation was examined under both restricted- and ample-oxygen supply conditions in a single fed-batch fermentation. Recombinant Escherichia coli transformed with the PHB production plasmid pSYLl07 was grown to reach high cell density (227 g/l dry cell weight) with a high PHB content (78% of dry cell weight), using a glucose-based minimal medium. A simple flux model containing 12 fluxes was developed and applied to the fermentation data. A superior closure (95%) of the carbon mass balance was achieved. When the data were put into use, the results demonstrated a surprisingly large excretion of formate and lactate. Even though periods of severe oxygen limitation coincided with rapid acetate and lactate excretion, PHB productivity and carbon utilization efficiency were not significantly impaired. These results are very positive in reducing oxygen demand in an industrial PHA fermentation without sacrificing its PHA productivity, thereby reducing overall production costs.

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