• Title/Summary/Keyword: 플라스미드

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Expression of recombinant plasmids harboring glucoamylase gene STA in saccharomyces cerevisiae (Glucoamylase 유전자 STA를 포함한 재조합 플라스미드들의 saccharomyces cerevisiae에서의 발현)

  • 박장서;박용준;이영호;강현삼;백운화
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.181-187
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    • 1990
  • STA gene coding glucoamylase was introduced into haploid Saccharomyces cerevisiae SHY3 and polyploid Saccharomyces cerevisiae 54. We constructed the recombinant plasmid by substituting the promoter region of alcohol dehydrogenase isoenzyme I gene for that of STA gene to increase the expression of STA gene and found that the activity of glucoamylase was increased in transformants. The plasmid stability was improved remarkably when we got the STA gene into the plasmid which had centromere. The activity of glucoamylase and transformation frequency of it, however, was decreased because of low copy number. Industrial polyploid strain was transformed with the recombinant plasmid having the $2\mu$ origin of replication and STA gene. It produced more alcohol than host when fermented in liquefied starch media. The industrial strain, however, was not transformed with the autonomously replicating plasmid containing centromere.

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Octane Biodegradability by Crude Oil4 tilizing Bacteria Carrying OCT Plasmid (OCT 플라스미드를 갖는 원유 분해세균에 의한 Octane 분해능)

  • 최순영;김창숙;황문옥;민경희;이명혜
    • Microbiology and Biotechnology Letters
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    • v.19 no.1
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    • pp.82-87
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    • 1991
  • Xanthomonns curnpestris M12, Xunthornonas sp. M28, Acinetuhucter Iwofz GI, and Klebsiella pneumoniae L25, Pseudomonas rnaltophiliu N246 were screened to increase the ability for crude oil utilization. All of these could utilize hexadecane and octane with the exception of N246 strain for only octane biodegradation. Thus N246, M12, and M28, strains were specially examined for octane oxidation. Octane biodegradation by three strains showed the optimal conditions at $30^{\circ}C$, pH 7.0~9.0, and 0.2~0.3% octane concentration as a substrate. It was found that P. multofihila N246 and X. curnpestns M12 had plasmid and the cured plasmid from N246 strain lost octane uitilization. Therefore, it was confirmed that certain genes for octane utilization were Iocated on OCT plasmid in N246 strain. The size of OCT plasmid in N246 strain was 118 kb. The N246 strain was resistant to ampicillin.

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Host Construction by Curing the Octopine Type Ti and Cryptic Plasmids in Agrobacterium tumefaciens KU12 (Agrobacterium tumefaciens KU12로부터 Octopine형 Ti 및 잠재 플라스미드의 제거에 의한 숙주 개발)

  • Ha, Un-Hwan;Lee, Yong-Woog;Moon, Hye-Yeon;Sim, Woong-Seop
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.53-59
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    • 1994
  • Agrobacterium tumefaciens KU12 contains pTiKU12 (240kb) of the octopine type Ti plamsid and pTi12 (45 kb) of the cryptic plasmid. To make the avirulent A. tumefaciens, the octopine type Ti plasmid, pTiKU12, was cured with elevated temperature (37${\circ}C$) and ethidium bromide (EtBr), respectively. Also the cryptic plasmid, pTi12, was cured by the introduction of recombinant plasmid, pYWXP, made by pTi12 replication origin and pUC19. pYWXP was cured by elevated temperature (37${\circ}C$) and EtBr simultaneously.

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Transfer and genetic recombination of antibiotic resistance genes occurring in water environment (수질환경에서 일어나는 항생물질 내성유전자의 전이와 재조합)

  • 김치경;이성기;김영창
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.245-250
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    • 1986
  • Antibiotic resistant bacteria isolated from the river water in Cheongju City were studied on the transfer of their R-plasmids in water environment. The R-plasmids were transferred by conjugation between the isolates ai the frequencies of 1.1$\times$10$^{-6}$ to 1.2$\times$10$^{-7}$ under laboratory condition and 1.2$\times$10$^{-7}$ to 1.0$\times$10$^{-9}$ in natural environment. The R-plasmids isolated from those bacteria were also transferred into the recipient cells of E. coli HB101 at the frequencies of 1.7-6.7$\times$10$^{-6}$. The AP$^{r}$Cm$^{r}$Tc$^{r}$-plasmid of isolate T-44 which were transformation by conjugation and transformation was determined to be 9.01 kilobses in molecular size. When the AP$^{r}$Cm$^{r}$Tc$^{r}$-plasmid DNA was treated with restriction endonuclease, the plasmid appeared to have one restriction site for EroRI and BamHI, respectively, and three sites for Pst I endonuclease.

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Identification of Octopine Type Ti Plasmid in Agrobacterium tumefaciens KU12 (Agrobacterium tumefaciens KU12내에 존재하는 Octopine Type Ti Plasmid의 확인)

  • 이용욱;음진성;심웅섭
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.292-299
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    • 1993
  • Agrobacterium tumefaciens KU12 isolated from Korea is able to induce tumors on various plants and catabolize octopine as a sole carbon and nitrogen source. A, tumefaciens KU12 contains three plasmids. Their sizes are 45.5 kb. 240 kb. and > 240 kb. respectively. For the purpose of identification of octopine type Ti plasmid, avirulent A, tumefacients A136 is transformed with plasmids isolated from KU12 by direct transformation. Transformants containing Ti plasmid were grown on AB medium containing octopine as a sole nitrogen source. The isolated strain, named KU911, contains only 240 kb plasmid. As a result of induction of crown gall and Southern hybridization with other octopine Ti plasmid pTiAch5, 240 kb plasmid named pTiKU12 was Ti plasmid.

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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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Complete genome sequence of multidrug-resistant Moraxella osloensis NP7 with multiple plasmids isolated from human skin (사람의 피부에서 분리한 다약제 내성이며 다수의 플라스미드를 갖는 Moraxella osloensis NP7 균주의 유전체 서열 분석)

  • Ganzorig, Munkhtsatsral;Lim, Jae Yun;Hwang, Ingyu;Lee, Kyoung
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.286-288
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    • 2018
  • Moraxella osloensis NP7 was isolated from human skin of a collage male and showed resistance to ${\beta}-lactam$ and aminoglycoside antibiotics. Herein, we report the complete whole-genome sequence and gene annotations of M. osloensis NP7. It possesses single circular chromosome and seven plasmids. Chromosome is 2,389,582 bp in length with the G + C content of 43.9% and encodes 2,065 protein-coding genes. The combined seven plasmids are 654,202 bp in size with the average G + C content of 40.5% and code for a total of 667 protein-coding genes. The chromosome of NP7 strain contains four ribosomal RNA operon copies, one transfer-messenger RNA gene, forty-seven tRNA genes, three riboswitch genes and three CRISPR arrays. Additional CRISPR array is found in the plasmid pNP7-1. The genes conferring resistance to ${\beta}-lactam$ and aminoglycoside antibiotics were predicted to reside in the plasmid pNP7-1.

DNA Rearrangement of TOL Plasmid in Pseudomonas putida PpGl Harbouring CAM Plasmid (CAM 플라스미드를 함유하는 Pseudomonas putida PpG1에서 TOL 플라스미드이 DNA 재배열)

  • 전효곤;조경연;고영희
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.433-436
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    • 1990
  • The TOL plasmid, pWWO, conjugally transferred from Pseudomonas putida mt-2 was dissociated into TOL* and TOL $\Delta$A in P. putidu PpGl carrying CAM plasmid. The TOL* was integrated into the CAM plasmid, and the resulting plasmid was designated as CAM::TOL*. The introduction of NAH plasmid, belonging to Inc P9 incompatibility group, into P. putida CSTBA carrying CAM::TOLt plasmid and TOL A plasmid did not affect m-toluate catabolism, but resulted in expelling the TOL $\Delta$ plasmid.

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Studies on Antimicrobial Susceptibility and Characteristics of R-plasmids and Antigens of High-level Gentamicin Resistant Enterococcus faecalis (Gentamicin 고도내성 Enterococcus faecalis균주의 항균제감수성, R-플라스미드 및 항원의 특성연구)

  • Kang, Hyun
    • Biomedical Science Letters
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    • v.1 no.1
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    • pp.55-72
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    • 1995
  • Forty gentamicin-resistant isolates of Enterococcus faecalis were selected from various clinical materials, determined their antimicrobial susceptibility, and studied there R-plasmid characteristics and polypeptide patterns. All of the isolates were susceptible to vancomycin. The MICs($\mu$/ml) of antimicrobial agents to the isolates were as follows; the MIC of gentamicin was 128 and $\geq$2040, ampicillin 1 and 1, chlorarmphenicol 2 and 8, erythromycin 32 and 256, and vancomycin 1 and 2. E. faecalis HL-1 strain had 8 plasmid DNA elements, HL-2 and HL-3 strains had 6, HL-4 had 7, HL-5 had 4, and HL-6 had 5. The 51.7 Kb of gentamicin resistance plasmid DNA was conjugally transferred from two strains of E. faecalis HL-1 and HL-6 to S. aureus SK 982. The plasmid transfer frequency between S. aureus SK 982 and E. faecalis HL-1 or E. faecalis HL-6 was 6.3$\times10^{-4} and 3.7$\times10^{-5}$, respectively. Plasmid curing ratio after the treatment of ethidium bromide(10$\mu$/ml) to E. faecalis tarnsconjugants R-1 and R-6 were about 51% and 67%, respectively. The tetracycline gene was located in 2.15 Kb plasmid of E. faecalis HL-1, but it was not found in the E. faecalis HL-6 by Southern blot analyses. The antigenic components of E. faecalis HL-1, HL-6, R-1 and R-6 strains were analyzed by SDS-PAGE and immunoblotting. The E. faecalis strains had 7 to 16 polypeptide bands, however their major proteins were 97.8 and 26.8 Kd. At the Immunoblotting, 97.8, 95.8, 74.8, 63.5, 33.7 and 26.8 Kd polypeptides of the strains showed major antigenic activities with patient's sera infected intra-abdominally with an E. faecalis strain.

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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.