• 제목/요약/키워드: Mutant screening

검색결과 134건 처리시간 0.028초

Gas Chromatographic Profiling법을 이용한 Tridecanedioic Acid를 생산해내는 Candida tropicalis Mutant의 탐색연구 (Gas Chromatographic Profiling for the Screening of Candida tropicalis Mutant Producing Tridecanedioic Acid)

  • 김정한;이상준;박형국;김경례
    • 한국미생물·생명공학회지
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    • 제19권2호
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    • pp.135-139
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    • 1991
  • Tridecanedioic acid (DC-13), starting material of the valuable musk ethylene brassylate, was obtained from n-tridecane by the Candida tropicalis mutant. The mutants were first obtained from primary screening step using the selective medium and then solid phase extraction sampling method was used for the selective isolation of organic acids from the cultured media of mutants. The resulting acids were directly converted to volatile tert-butyldimethyl silyl delivatives, which were then analyzed by gas chromatography. The efficient GC profiling method was used for the rapid identification of the mutant producing DC-13 in large quantity, and for the optimization of the culture conditions of mutant. The optimal culture conditions were found as follows: pH 8.0, 30$^{\circ}C$, 250rpm, 48hour of culture and $(NH_4)_2HPO_4$ as nitrogen source.

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The screening & characterization of super-producing recombinant Hansenula polymorpha mutant

  • 강환구;박형수;이충열;유병일;유은정;이선;황선덕;강현아;이상기
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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    • pp.684-687
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    • 2000
  • The super-producing recombinant H.polymorpha mutant is obtained by double membrane screening technique combined with optimum mutation method. The characterization of mutant is carried out to find the change of mutant in m-RNA level, cell wall leakage, protease level and methanol utilization metabolic flux. The change of these properties of mutant was figured out.

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Cephalosporin C 생물전환을 위한 Trigonopsis variabilis (ATCC10679) 변이균주의 간편한 선별 및 D-amino acid oxidase 유전자 배열 (Rapid Screening of Mutant Strains of Trigonopsis variabilis (ATCC10679) for Cephalosporin C Bioconversion and Sequences of D-amino acid oxidase Genes)

  • 강용호;박선영
    • KSBB Journal
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    • 제14권2호
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    • pp.235-240
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    • 1999
  • Trigonopsis variabilis ATCC10679 (TW)의 변이균주를 선별하기 위하여 쉽고 간편한 균주 선별방법을 개발하였다. 변이 균주(T26)의 D-Amino acid oxidase (D-AAO)는 야생균주(TW)의 D-AAO 효소보다 cephalosporin C에 대한 기질특이성이 약 30% 높았다. 이들 두 균주에서 D-AAO 유전자를 클로닝하여 각 유전자를 비교해본 결과 811번 우치의 T가 C로 변경되었으며 이에 따른 아미노산은 Phe-258이 Ser-258로 바뀌었다.

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Mutagenic Analysis of hPNMT Confirms the Importance of Lys57 and the Inhibitor Binding Site

  • Jeong, Ki-Woong;Kang, Dong-Il;Lee, Jee-Young;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • 제32권2호
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    • pp.455-458
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    • 2011
  • In previous report, with the aid of receptor-oriented pharmacophore-based in silico screening, we characterized three novel hPNMT inhibitors (YPN010, YPN016, and YPN017) and proposed that the hydrogen bonding interaction between inhibitors and side chain of Lys57 is very important to inhibitory activity of hPNMT. To confirm the importance of Lys57, mutant with substitution of Lys57 with Ala was cloned and binding study was performed for a K57A mutant of hPNMT using STD-NMR and fluorescence experiments. The binding constants for three novel inhibitors with mutant hPNMT were dramatically decreased compared to those with wild-type protein. K57A mutant-induced conversion of noradrenaline to adrenaline was suppressed about 95 % compared to wild-type hPNMT. Mutagenic analysis using a K57A mutant confirmed the importance of the Lys57 residue in binding of the inhibitor candidate to hPNMT as well as enzymatic activity of hPNMT, implying that these results are consistent with our binding model.

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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Isolation of Deletion Mutants by Reverse Genetics in Caenorhabditis elegans

  • Park, Byung-Jae;Lee, Jin ll;Lee, Jiyeon;Kim, Sunja;Choi, Kyu Yeong;Park, Chul-Seung;Ahn, Joohong
    • Animal cells and systems
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    • 제5권1호
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    • pp.65-69
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    • 2001
  • Obtaining mutant animals is important for studying the function of a particular gene. A chemical mutagenesis was first carried out to generate mutations in C. elegans. In this study, we used ultraviolet-activated 4,5',8-trimethylpsoralen to induce small deletion mutations. A library of mutagenized worms was prepared for recovery of candidate animals and stored at $15^{\circ}C$ during screening instead of being made into a frozen stock library. In order to isolate deletion mutations in target genes, a polymerase chain reaction (PCR)-based screening method was used. As a result, two independent mutants with deletions of approximately 1.0 kb and 1.3 kb were isolated. This modified and improved reverse genetic approach was proven to be effective and practical for isolating mutant animals to study gene function at the organismal level.

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Aspergillus flavus의 강력 protease생성 돌연변이의 유발 (On a highly proteolytic mutant strain of Aspergillus flavus)

  • 이영녹;박용근;고상균
    • 미생물학회지
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    • 제18권2호
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    • pp.51-58
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    • 1980
  • Mutational experiments were performed to improved to improve the protease productivity of Aspergillus flavus KU 153, which is selected among the wild strains. A UV-induced mutant strain having high protease productivity was obtained by the use of the clear zone method as a simple criterion for a primary screening test. Neutral and alkaline protease activities of hte mutant strain were higher than 1.8 times, comopared with those of the parental strain, respectively, while in the case of acid protease, it was 2.7 times. The mutant strain selected was more powerful in the production of cellulase and amylase, as well s protease in wheat bran, compared with those of the parental strain. protease production of the parental strain has reached maximum level at 3 days culture, while alkaline nad neutral protease production of the mutantstrain has reached at 2 days culture. On the other hand, the mutant strain formed the spore slowly, compared with the parental strain. Column chromatography of the neutral protease on DEAE-Sephadex A-50 showed that the mutant strain was not induced the formation of another neutral protease isozyme, but induced the variation in the function of regulatory gene.

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