• Title/Summary/Keyword: Transformant

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Antimutagenic Effects on Aflatoxin $B_1$ of Soybean Pastes Fermented by Bacillus Strains

  • Lee, Eun-Ju;Lee, Hyo-Jin;Kim, Jong-Kyu
    • Food Science and Biotechnology
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    • v.14 no.6
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    • pp.878-880
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    • 2005
  • Antimutagenic effects of methanol extracts of various soybean pastes against aflatoxin B1 were examined using Salmonella typhimurium TA98 and TA100. Antimutagenic activities of boiled soybeans, Japanese Miso, traditional Korean soybean pastes, soybean pastes fermented by wild type strains, and soybean pastes fermented by mutants, transformants, and cell fusants were 53.6 to 54.6%, 73 to 79.7%, 78.3 to 95.7%, 85 to 97.1%, 71.9 to 78.3%, 65.5 to 77.7%, and 73.4 to 79.0%, respectively. Soybean pastes fermented by wild type strains showed higher activities than those fermented by mutant, transformant, and cell fusant strains.

Molecular Cloning and Expression of the Acetyl Xylan Esterase Gene of Bacillus stearothermophilus in Escherichia coli (Bacillus stearothermophilus Acetyl Xylan Esterase 유전자의 크로닝과 Escherichia coli에서의 발현)

  • 김인숙;조쌍구;최용진
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.542-548
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    • 1993
  • Bacillus stearothermophilus was shown to express multiple xylanolytic enzymes including acetyl xylan esterase. Genomic DNA of the strain partially digested with HindIII was ligated into the HindIII site of pBR322, and expressed in E. coli HB101 cells in order to clone the gene for acetyl xylan esterase. One transformant among 4000 screened formed a clear zone around its colony on the LB agar supplemented with 1.0% tributyrin. The functional clone harbored the recombinant plasmid pKMG5 with an insert of 5.1kb.

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Introduction and Expression of the Urease Gene in Mosquitocidal Bacillus sphaericus 1593 (세균성 Urease Gene에 의한 모기유충 방제균 Bacillus sphaericus 1593의 형질전환)

  • 한길환;김상달
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.390-396
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    • 1995
  • Bacillus sphaericus 1593 is a larvicidal toxin-producing mosquitocidal bacterium. The toxin contains a parasporal crystalline inclusion which is composed of a protein that is activated under alkaline condition. To enhance alkaline environment around toxin protein, cryptic plasmid cured, B. sphaericus 1593 was transformed by the Bacillus pasteurii urease gene which generate ammonia from urea. Transformant produced urease at about 80% more than wild type strain. B. sphaericus 1593, and the urease gene was stably maintained. It also produced crystalline toxin protein at the same level as the wild type strain B. sphaericus 1593.

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Expression of the Glyphosate Resistant Gene, cp4-epsps, through Plastid Transformation in Rice (Oryza sativa L.) (벼 색소체 형질전환을 이용한 글리포세이트 저항성 유전자 cp4-epsps의 발현)

  • Kang, Kyung-Su;Kim, Min-Kyun
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.75-84
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    • 2006
  • Heteroplasmic rice plastid transformant was generated using suspension cells as bombardment materials. PCR analyses confirmed incorporation of aadA and cp4-epsps genes into the rice plastid genome by homologous recombination events via the flanking sequences of the trnI and trnA. Transplastomic calli were actively proliferated when cultured on AAM2 medium supplemented with various concentrations (500-3000 mg/L) of streptomycin in dark condition, and transplastomic suspension cells showed resistance to nonselective herbicide, glyphosate. Through 'agarose pie selection' method, heteroplastomic calli, containing considerably high level of transplastome and expressing the CP4 EPSPS protein, were obtained. They were further regenerated to green shoots with healthy roots.

Characterization of Heat-Stable Enterotoxin of Enterotoxigenic Escherichia coli eKT-53 (장독성 대장균 eKT-53균주의 내열성 장독소의 성질)

  • 도대홍;김교창;김도영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.6
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    • pp.621-628
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    • 1991
  • Heat-stable enterotoxin(ST) from enterotoxigenic E. coli eKT-53($ST^{+}\;LT^{-}$, transformant from isolate KM-7) that was produced in succinate salts medium. The culture supernatant(crude ST) was purifed by mulitpled steps and investigated some characterization of the ST. The heatstability of purified ST activity was completely lost by treating at $100^{\circ}C$ for 30minutes. ST activity was lost by treatment at pH 1 and 12 conditions, while the activity was not reduced by treatment at pH 2~10, and then the ${\alpha}-amylase$ and pepsin was not decreased activity but disulfide reducing agnets was lost the activity. The molecular weight of the purified ST was approximately 4,200, the isoelectric point was about 4.0.

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Molecular cloning of phospho-$\beta$-galactosidase gene of lactobacillus casei in escherichia coli (Lactobacillus casei의 phospho-$\beta$-galactosidase 유전자의 대장균내 분자클로닝)

  • 문경희;박정희;최순영;이유미;김태한;김연수;민경희
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.188-193
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    • 1989
  • Gene for lactose catabolism in Lactobacillus casei SW-M1 was encoded by a 60Kb metabolic plasmid. A derivative of only 10kb, pPlac 15 of recombinant plasmid, was constructed by introducing into pBR322 and was cloned into E. coli using restriction endonuclease Pst I. A 10kb insery DNA in plasmid pBR322 was identified as a gene encoded phospho-$\beta$-galactosidase by the determination of enzyme activity. Phospho-$\beta$-galactosidase was apparently expressed in E. coli. The enzyme activities of cell-free extract from transformant E. coli HB101 carrying pPLac 15 DNA were not different from that of L. casei as a donor strain on the basis of enzyme properites. However, specific activity of phospho-$\beta$-galactosidase in the cloned strain with Lac $Y^{-}$ phenotype of E. coli HB101 was lower than that in L. casei strain.

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폐염균의 형질전환 및 recP유전자의 클로닝

  • 이동권
    • The Microorganisms and Industry
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    • v.15 no.2
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    • pp.24-28
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    • 1989
  • 폐염균의 특징은 폐염, 류마티스성 심장병, 수막염 등의 질병을 일으키는 병원균이라는 것과 아주 높은 효율로 형질전환을 할 수 있다는 사실이다 ($10^{8}$ 개의 균에 염색체 DNA 1.mu.g을 써서 실험하였을 때 $10^{6}$개의 transformant를 얻을 수 있음). 폐염균이 자연상태에서 이렇게 높은 효율의 형질전환을 할 수 있는 이유는 정상생리의 일부로서 형질전환에 관계된 유전자가 형질전환을 할때 발현되기 때문이다. 그러나 폐염균 형질전환에 대한 연구는 아직도 초기단계라고 할 수 있으며 그 이유는 현재 전세계적으로 극소수의 연구자들만이 연구를 수행하고 있기 때문이다. 즉, 미국 Brookhaven National Laboratory의 Lacks와 프랑스 CNRS의 Claverys는 mismatch repair에 대한 연구를, Illinois 대학의 Morrison은 competence에 대해, 미국 Rockefeller 대학의 Hotchkiss와 Tomasz는 DNA binding을 연구하고 있을 뿐이다. 본 논고에서는 폐염균 형질전환에 대해 지금까지의 연구 상태와 문제점, recP의 클로닝에 대해 소개하고자 한다.

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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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Expression of Proteinase Inhibitor II gene in Transgenic Flowering Cabbage, Brassica oleracea var. acephala DC. (형질전환된 꽃양배추에서 Proteinase Inhibitor II 유전자의 발현)

  • 김창길;정재동
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.95-98
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    • 1998
  • Hypocotyl explants of flowering cabbage were cocultured with Agrobacterium tumefaciens LBA4404;;pGA875 harboring proteinase inhibitor II(PI-II) cDNA and then regenerated into plants. Sucessful transcripts of PI-II gene were detected by RNA dot blot analysis. Bioassay was conducted on transgenic flowering cabbage. It was confirmed that insecticidal activities of transformants were much higer than that of control plants. In progeny test of hansformants, 27.4% of T$_1$ seeds was resistant on MS medium containing 20 mg/L kanamycin.

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Gene Expression and Iron Accumulation in Progeny of Transformants Introduced Fp1 Gene Encoding the Iron Storage Protein in Red Pepper (Capsicum annuum L.)

  • Kang, Kwon-Kyoo;Kim, Young-Ho
    • Plant Resources
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    • v.4 no.1
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    • pp.26-30
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    • 2001
  • To improve the iron content of red pepper, we have transferred the entire coding sequence of the ferritin gene(Fpl) into Capsicum annuum (L. cv. Chungyang and Bukang) by Agrobacterium mediated transformation. Transformants were found to contain the Fp1 gene at up to three loci, increased distinct iron content changes. In transgenic plants, iron content was as much as 7-fold to 8-folds greater than that of their untransformed counterparts. Furthermore, the Rl progenies from transformant(A7, A8) co-segregated into a 15:1 ratio for both Kanamycin resistance and genotype of high iron.

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