• Title/Summary/Keyword: UV-mutagenesis

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Trp, Thr Analogue 복합 저항성 Saccharomyces cerevisiae 균주 개발 (Isolation of Trp, Thr Overproducing Strain of Saccharomyces cerevisiae)

  • 염형준;이승현;김선혜;선남규;안길환;이봉덕;원미선;송경빈
    • 한국식품영양과학회지
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    • 제33권6호
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    • pp.1017-1021
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    • 2004
  • Thr, Trp을 overproduce하는 효모 균주를 분리하기 위하여 Sacchromyces cerevisiae를 UV와 EMS로 mutagenesis한 후 screen하였다. Thr analogue인 hydroxynorvaline이 UV mutagenesis 후 Thr overproducing 하기 위해 사용되었다. 31 mutant 중 아미노산 분석 결과에 의해 TC 5-1이 선정되었고 다시 Trp overproducing 위해 EMS mutagenesis 하였다. 8개의 mutant가 flurotryptophan을 이용하여 Thr, Trp을 overproduce하는 mutant로 선정되었다. 아미노산결과에 의해 그 중 TC 6-1이 최종 효모 균주로 선정되었다.

Aspergillus nidulans에서 MNNG 선 처리시의 생존도와 돌연변이 유발에 대한 Adaptive response 및 Cell stage 따른 UV와 MNNG에 대한 치사율 조사 (Adaptive Responses on Survival and Mutagenesis during MNNG Pretreatmeat and Lethality to UV MNNG at Different Cell Stages in Aspergillus nidulans)

  • 채순기
    • 자연과학논문집
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    • 제9권1호
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    • pp.45-52
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    • 1997
  • 저농도의 MNNG가 Aspergillus nidulans의 생존도 및 돌연변이 유발에 끼치는 영향을 조사하였다. Nontoxic하고 submutagenic한 농도의 MNNG 선 처리는 높은 농도로 처리 시의 치사율 및 돌연변이 유발을 낮추지 못했다. 이러한 결과는 Aspergillus nidulans에는 MNNG 에 의한 adaptive response가 일어나지 않는다는 것을 시사하고 있다. 발아 과정의 첫 번째 체세포 분열에서, 시간별로 MNNG에 대한 치사율을 조사하고 UV에 의한 생존도와 비교하였다. UV나 MNNG 처리 시 치사율은 S 세포 시기 직전까지 증가하였다가, DNA 복제 시에는 감소함을 나타내었다. MNNG 처리 시는 UV와 달리 G2세포시기에 치사율이 가장 높았다.

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Gene-Specific Repair of 6-4 Photoproducts in Trichothiodystrophy Cells

  • Nathan, Sheila;Van Hoffen, Anneke;Mullenders, Leon H.F.;Mayne, Lynne V.
    • BMB Reports
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    • 제32권6호
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    • pp.554-560
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    • 1999
  • TTD1BI cells are non-hypersensitive to UV irradiation and perform normal genome repair of pyrimidine dimers but fail to excise 6-4 photoproducts and, concomitantly, are unable to restore RNA synthesis levels following UV irradiation. This pointed to a detect in gene-specific repair and this study was undertaken to examine repair of 6-4 photoproducts at the gene-level. The results indicated a defect in gene-specific repair of 6-4 photoproducts in active genes, although strand-specificity of 6-4 photoproduct removal was essentially similar to that of normal cells. These findings indicate that the near normal UV resistance of TTD1BI cells may be due to the inability of these cells to remove DNA lesions preferentially, as well as to the cells opting out of the cell cycle to repair damage before resuming replication.

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Biosorption of uranium by Bacillus sp.FB12 isolated from the vicinity of a power plant

  • Xu, Xiaoping;He, Shengbin;Wang, Zhenshou;Zhou, Yang;Lan, Jing
    • Advances in environmental research
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    • 제2권3호
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    • pp.245-260
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    • 2013
  • Biosorption represents a technological innovation as well as a cost effective excellent remediation technology for cleaning up radionuclides from aqueous environment. In the present study, a bacteria strain FB12 with high adsorption rate of uranium ion was isolated from the vicinity of the nuclear power plant. It was tentatively identified as Bacillus sp.FB12 according to the 16S rDNA sequencing. Efforts were made to further improve the adsorption rate and genetic stability by UV irradiation and UV-LiCl cooperative mutagenesis. The improved strain named Bacillus sp.UV32 obtains excellent genetic stability and a high adsorption rate of 95.9%. The adsorption of uranium U (VI) by Bacillus sp.UV32 from aqueous solution was examined as a function of metal ion concentration, cell concentration, adsorption time, pH, temperature, and the presence of some foreign ions. The adsorption process of U (VI) was found to follow the pseudo-second-order kinetic equation. The adsorption isotherm study indicated that it preferably followed the Langmuir adsorption isotherm. The thermodynamic parameters values calculated clearly indicated that the adsorption process was feasible, spontaneous and endothermic in nature. These properties show that Bacillus sp.UV32 has potential application in the removal of uranium (VI) from the radioactive wastewater.

알파-아밀라제 저해제 생산 방선균 Streptomyces minoensis DMCJ-144의 균주개량 (Strain Improvement of Streptomyces minoenisis DMCJ-144, An ${\alpha}$-Amylase Inhibitor Producing Actinomycetes)

  • 최응칠;김숙경;강동희;이재우;김병각
    • Biomolecules & Therapeutics
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    • 제1권1호
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    • pp.26-30
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    • 1993
  • Strain of treptomyces minoensis DMCJ-144 was tried to be improved so that it produces much more the $\alpha$-amylase inhibitor. Streptomyces minoensis DMCJ-144 was treated with 1 mg/mι (pH 9.0) of N-methyl-N'-nitro-N-nitrosoguanidine at $30^{\circ}C$ for 60 min and irradiated with UV light distanced 30 cm for 20 min. After mutagenesis, surviving colonies were cultured on the CM contaning acriflavine ($10{\mu}g/ml$) three times in order to enhance the mutability. And then through multi-level screening, colonies that ${\alpha}$-amylase inhibitor productibility. was Improved were selected by modified-blue value method. After third acriflavine treatment, $\alpha$-amylase inhibitory activities of selected colonies were found to be much better as compared with that of parent strain. One mutant strain showed 5.4 time inhibitory activity than the parent strain.

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Strain Improvement of Candida tropicalis for the Production of Xylitol: Biochemical and Physiological Characterization of Wild-type and Mutant Strain CT-OMV5

  • Rao Ravella Sreenivas;Jyothi Cherukuri Pavanna;Prakasham Reddy Shetty;Rao Chaganti Subba;Sarma Ponnupalli Nageshwara;Rao Linga Venkateswar
    • Journal of Microbiology
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    • 제44권1호
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    • pp.113-120
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    • 2006
  • Candida tropicalis was treated with ultraviolet (UV) rays, and the mutants obtained were screened for xylitol production. One of the mutants, the UV1 produced 0.81g of xylitol per gram of xylose. This was further mutated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and the mutants obtained were screened for xylitol production. One of the mutants (CT-OMV5) produced 0.85g/g of xylitol from xylose. Xylitol production improved to 0.87 g/g of xylose with this strain when the production medium was supplemented with urea. The CT-OMV5 mutant strain differs by 12 tests when compared to the wild-type Candida tropicalis strain. The XR activity was higher in mutant CT-OMV5. The distinct difference between the mutant and wild-type strain is the presence of numerous chlamvdospores in the mutant. In this investigation, we have demonstrated that mutagenesis was successful in generating a superior xylitol-producing strain, CT-OMV5, and uncovered distinctive biochemical and physiological characteristics of the wild-type and mutant strain, CT-OMV5.

Combination Strategy to Increase Cyclosporin A Productivity by Tolypocladium niveum Using Random Mutagenesis and Protoplast Transformation

  • Lee, Mi-Jin;Duong, Cae Thi Phung;Han, Kyu-Boem;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • 제19권9호
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    • pp.869-872
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    • 2009
  • The cyclic undecapeptide cyclosporin A (CyA), one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. To increase CyA productivity by wild-type T. niveum (ATCC 34921), random mutagenesis was first performed using an antifungal agar-plug colony assay (APCA) selection approach. This generated a mutant strain producing more than 9-fold greater CyA than the wild-type strain. Additionally, a foreign bacterial gene, Vitreoscilla hemoglobin gene (VHb), was transformed via protoplast regeneration and its transcription was confirmed by RT-PCR in the UV-irradiated mutant cell. This led to an additional 33.5% increase of CyA production. Although most protoplast-regenerated T. niveum transformants tend to lose CyA productivity, the optimized combination of random mutagenesis and protoplast transformation described here should be an efficient strategy to generate a commercially valuable, yet metabolite low-producing, fungal species, such as CyA-producing T. niveum.

Screening of Phytase Overproducing Strains in Aspergillus spp. by UV Mutagenesis

  • Lee, Eung-Suek;Paik, In-Kee;Hahm, Young-Tae
    • Mycobiology
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    • 제28권3호
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    • pp.119-122
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    • 2000
  • Phytases (myo-inositol hexakisphosphate phosphohydrolase; EC 3.1.3.8) are enzymes which catalyze the hydrolisys of phytate into myo-inositol and inorganic phosphates. Phytases are found in plants and a variety of microorganisms. Aspergillus species were treated with 254 nm of UV irradiation for the screening of phytase overproducing mutant strains. At 15 minute irradiation, the survivals of population were less than 5%, and UV irradiation time was decided at 20 minute for the isolation of mutant strains. Four UV mutant strains in A. oryzae (YUV-47, -169, -341, -511) and six in A. ficuum (FUV-17, -36, -69, -193, -317, -419) were isolated on PSM media containing ammonium phosphate. The specific enzyme activities of A. ficuum mutants are 110 to 140% higher than that of wild type.

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돌연변이와 DNA 손상회복에 미치는 muc 유전자의 기능 (Function of muc Gene on Mutagenesis and DNA Repair)

  • 전홍기;이상률;백형석
    • 미생물학회지
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    • 제28권3호
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    • pp.192-198
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    • 1990
  • 플라스미드 pKM101과 이의 돌연변이제 pSLA의 mutator 유전자를 subcloning하여 재조합 플라스미드 pJB200과 pJB210을 선별하였고, umuC36- uvrA6-(TK 610) 균주에 도입하여 UV와 MMS에 대해서 보호효과와 돌연변이율에 미치는 영향을 조사하였다. 재조합된 플라스마드는 UV와 MMS에 대해서 nonmutability인 umu- 균주에서 완전히 돌연변이율을 회복시켰고 보호효과를 크게 증가시켰다. 이 사실은 muc 유전자가 cloning된 재조합 플라스미드가 돌연변이원의 처리에 의해 효과적인 발현을 하며, muc 유전자 만으로도 pKM101의 기능을 나타낸다는 것을 확인하였고, pSLA의 muc 유전자가 그 효과에 었어서 높은 영향을 미쳤다. 또한 pKM101의 muc gene을 포함한 pJB210 은 recA - (JC2926) 균주에서 돌연변이 유발능에 영향을 미치지 못한 것은 muc 유전자가 recA 유전자에 의존함을 알 수 있었다.

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섬유소분해효소 생산증진을 위한 Penicillium verruculosum의 균주개량 (Strain Improvement of Penicillium verruculosum for High Cellulase Production by Induced Mutation)

  • 정기철
    • 한국미생물·생명공학회지
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    • 제15권6호
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    • pp.388-395
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    • 1987
  • 천연섬유소 분해활성이 우수하고 그 효소 유도기구도 Trichoderma reesei와는 다른 Penicillium verruculosum F-3을 모균주로 사용하여 돌연변이 처리에 의한 유전적 개량을 시도함으로써 cellulase 생산성이 증진된 조절변이주를 얻고자 변이주 유도조건 변이주에 의한 cellulase 생성조건을 검토하였다. 한천평판상에서의 변이주의 선택분리 효율을 향상시키기 위하여 각종 colony 소형화제의 영향을 검토한 바 Oxgall을 배지에 1.5% 첨가하였을 때가 가장 좋았다. Cellulase 고생산성 변이주 선정의 한 지표로서 대사산물 억제의 해제를 선택했다. P. verruculosum F-3은 glucose 또는 glycerol 농도 1% 이상에서 본 효소생성이 억제되었다. 변이주 유도조건으로서 UV조사의 경우는 19분 처리로 약 0.2%, NTG 처리의 경우는 200$\mu\textrm{g}$/$m\ell$ 농도로 1시간처리로 48%의 생존율을 나타냈다. 변이처리 한 균주를 여지붕괴도, cellulose agar plate에서의 clear zone의 크기, cellulose powder 배지로 배양한 조효소액의 여지분해활 성을 조사하여 우수균주로서 UV-9, UV-10 및 NTG-3을 최종 선발했다. 각종 탄소원을 함유하는 배지에서의 cellulase 생산성을 조사한 바 KC-M-W 배지로 배양한 UV-9, UV-10 및 NTG-3의 여지분해 활성은 친주보다 각각 34%, 55%, 41% 증가되었으나, UV-9 및 NTG-3은 COA 자화능이 현저히 저하되었다. 변이주 UV-10은 COA-4 배지로 배양했을 때 친주에 비해 단백질량 30%, Avicel 분해활성 30%, 여지분해활성 20%, salicin 분해활성 50% 증가가 인정되었고, 비록 역가는 낮았지만 glucose 및 cellobiose를 함유하는 배지에서 CMC 및 salicin 분해활성을 구성적으로 생산하였다.

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