• 제목/요약/키워드: Mutant

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비스판 변이주에 의한 활성아포 생산성 증가

  • 전경동;이광호;김원석;백현동
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.275-278
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    • 2000
  • 본 연구에서 비스판균에 비해 아포형성능이 우수한 변이주를 선별하고, 발효조 배양에서 활성아포 생산성의 증가를 확인하였다. 최종 선별된 변이주 KD21균주를 5 L jar fermenter에서 배양한 결과, 36시간만에 총균수는 $5.5{\times}10^9$ CFU/mL에 도달하고, 최대 활성아포수는 $2.7{\times}10^9$ CFU/mL에 도달하였다. 이는 비스판 친균주에 비해 활성아포의 생산성이 77%증가하였음 확인 할 수 있었다. 장질환 치료제로 판매되고 있는 비스판 제제의 활성아포 생산성이 우수한 변이주를 획득함으로서 친균주에 비해 가격경쟁력을 가질수 있을것으로 판단된다.

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Phycobilisome composition in Chondrus crispus (Gigartinales, Rhodophyta) from a wild type strain and its vegetatively derived green mutant

  • Cornish, M. Lynn;O' Leary, Stephen J.B.;Garbary, David J.
    • ALGAE
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    • 제28권1호
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    • pp.121-129
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    • 2013
  • Intact phycobilisomes from a wild-type red Chondrus crispus and its vegetatively derived green mutant were isolated by centrifugation through a discontinuous sucrose density gradient. Pigment composition was subsequently characterized by spectrophotometry. Vegetative thalli of the two strains grown together for six months in the laboratory resulted in different pigment profiles. Two pigmented phycobilisome bands appeared in the sucrose gradient of the wild-type alga, a purple coloured one, and a pink one, whereas only a single blue band appeared in the gradient of the green mutant. Spectrophotometric and fluorescence analyses identified the phycobiliprotein composition of the purple band as the typical phycoerythrin-phycocyanin-allophycocyanin complement in the wild-type, but there was no detectable phycoerythrin present in the blue band of the green mutant. Sodium dodecyl sulphate, preparative polyacrylamide gel electrophoresis analysis confirmed the presence of allophycocyanin subunits in all extracts, but firm evidence of an R-phycoerythrin linker polypeptide in the blue band was missing. These results highlight the ability of C. crispus to adapt to a phycoerythrin deficiency by adjusting light harvesting pigment ratios.

Construction of multiple mutant strains by mating procedures for the cloning of pmn and pmb genes encoding amino acid permeases in neurospora crassa

  • Han, Hyo-Young;Min, Kyung-Hee
    • Journal of Microbiology
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    • 제33권2호
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    • pp.142-145
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    • 1995
  • The pumb gene encoding a basic amino acid transport protein in Neurospora crassa could be cloned by using a mutant strain defective in pmb gene as a host strain, using a negative selection on the media containing amino acid analogue canavanine. To select positive transformants of the genes for cloning, an auxotrophic marker (his-2) was added to a pmb mutant strain by mating ; a triple mutant (pmn : pmb : his-2) was constructued by crossing a strain defective in basic amino acid transport system (# 1683-bat um 535 "A") to a double mutant strain defective in neutral amino acid transport and histidine production (mitrol : his-2 "a"). Crossing was performed on synthetic crossing (SC) media containing histidine. The pmn : pmb and pmn :pmb : his-2 strains were selected among the progeny colonies from crosses on plates containing 5- .mu.g/ml para-fluoro-phenylalanine (PFPA), 200 .mu.g/ml canavanine, and 500 .mu.g/ml histidine. The selected colonies were cultured on minimal media with or without histidine for discarding pmn : pmb strain, because the pmn : pmb : his -2 strain grows only on histidine containing media. The pmn :pmb : his-2 strain selected can be used as a host strain for the cloning of the pmb and the pmn genes from a Neurospora genomic library by means of positive selections.

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Riboflavine 생산성국균(生産性麴菌)에 의(依)한 Riboflavin 강화(强化)된장의 제조시험(製造試驗) (A study on manufacturing of Riboflavine fortified soybean mash with an exceedingly Riboflavine productive koji mold mutant)

  • 박태준;조덕현;김호식
    • Applied Biological Chemistry
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    • 제2권
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    • pp.17-21
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    • 1961
  • The manufacture of riboflavine fortified Dwen-Jang has been tried with an exceedingly riboflavine productive Aspergillus oryzae #612 mutant which has been developed by the authors. Both the rice and barley koji of this mutant and Aspergillus sojae have been prepared. Their riboflavine production, saccharifying and protease activities have begin compared The riboflavine fortified Dwen-Jang has been manufactured using the barley koji of riboflavine productive mutant. Their riboflavine content and qualities have been studied comparing with an ordinary Dwen-Jang which has been prepared with the barley kojo of A. sojae strain. The following results have been obtained. (1) The baley koji was superior in riboflavine production and protease activity, inferior in saccharifying ability than rice koji both with A. oryzae #612 and A. sojae. (2) In barley koji, the mutant, A. oryzae #612, produces 1.5 times riboflavine than A. sojae and shows stronger saccharifying and protease activities than the latter. (3) The riboflavine fortified Dwen-Jaug manufactured contained $5.2{\gamma}/g$ of riboflavine, about 1.5 times that of A. sojae. The higher contents of free sugar and free amjno nitrogen have been observed than the ordinary Dwen-Jang manufactured with A. sojae.

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Sll0396 regulates transcription of the phycocyanin genes in Synechocystis sp. PCC 6803

  • Oh, In-Hye;Kim, Ho-San;Chung, Young-Ho;Kim, Young-Hye;Park, Young-Mok
    • Plant Biotechnology Reports
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    • 제4권3호
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    • pp.193-199
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    • 2010
  • An olive-green mutant was generated in Synechocystis sp. strain PCC 6803 by inactivation of the sll0396 gene. Whole-cell absorption spectra of the mutant revealed the missing of phycocyanin peak. An investigation of the low-temperature fluorescence emission spectra revealed that the $sll0396{\Omega}$ mutant has a reduced amount of phycocyanin. Western blot analysis showed that the mutant contained less phycocyanin ${\beta}$- and ${\alpha}$-subunits and lacked the 30- and 32-kDa linker polypeptides, and northern blot analysis revealed that the transcription of the 1.4-kb cpcBA gene encoding the phycocyanin ${\beta}$- and ${\alpha}$-subunits was lower in the mutant. The Sll0396 protein has a DNA-binding motif and shares homology with known response regulators. Our results indicate that Sll0396 plays a regulatory role in the transcription of the phycocyanin genes during phycobilisome synthesis.

Growth, Nitrogen Metabolism, and Nodulation of Hypernodulating Soybean Mutant Affected by Soil Fertility

  • Ha, Bo-Keun;Lee, Suk-Ha
    • 한국작물학회지
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    • 제46권2호
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    • pp.145-149
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    • 2001
  • This study was performed to evaluate the growth and nodulation characters of hypernodulating soy-bean mutant, SS2-2, and to know the growth and yield performance of the mutant in infertile soil. Soil fertility was adjusted by mixing the different ratios of soil components including clay, river sand, and horticultural bed, which resulted in fertile and infertile soil. Dry weight, nitrogen concentration, and leaf nitrate reductase of each plant were measured around V6 stage (47 days after planting) and around R3 stage (82 days after planting). There were significant effects of soil fertility and soybean genotype on the total dry weights including root, nodule, stem, leaf, and pod dry weight at V6 and R3 stages. Total dry weight of hypernodulating mutant, SS2-2, was clearly less than that of its wild type, Sinpaldalkong 2. However, nodule development on the roots of SS2-2 was much greater than that of Sinpaldalkong 2, regardless of soil fertility. Though SS2-2 was smaller in plant size than Sinpaldalkong 2, genotypic difference in total nitrogen content was not significant at both V6 and R3 stages because SS2-2 fixed more nitrogen biologically than its wild type in the root nodule. The SS2-2 mutant showed lower plant yield in both infertile and fertile soil. The SS2-2 contained more crude seed protein than Sinpaldalkong 2, and was characterized with reduced top and root growth.

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Characterization of a Photosynthetic Mutant Selected by Increased Formation of Poly-3-Hydroxybutyrate in Rhodobacter sphaeroides

  • Lee, Il-Han;Kho, Dhong-Hyo;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • 제8권6호
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    • pp.714-718
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    • 1998
  • Various mutants either lacking or having decreased levels of light-harvesting complexes and reaction center complex were obtained with a high frequency by an increased formation of poly-3-hydroxybutyrate (PHB) in Rhodobacter sphaeroides. One of the photosynthesis-defective mutants, PY-17, which was devoid of any of the light-harvesting complexes (B800-850, B875) as well as the reaction center complex, was analyzed further. The mutant showed substantial transcription of the puhA, pufKBALMX, and pucBAC operons coding for the structural proteins of the photosynthetic complexes although each of the activities was lower than that of the wild type. Translation of the pufKBALMX and pucBAC operons were also active in the mutant although with activities different from the corresponding one of the wild type. From these results the mutation appears to exert its effect at the post-translational level of the photosynthetic complex assembly. Complementation of the photosynthesis-defective phenotype of the mutant was achieved with an about 12-kb DNA region containing the puhA gene. The relationship between the formation of PHB and photosynthetic complexes is discussed.

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Aminopeptidase M 저해제를 생산하는 Streptomyces sp. SL-387 (KCTC 0102BP) 변이주의 특성 (Characterization of the Mutant of Streptomyces sp. SL-387(KCTC 0102BP) Producing Aminopeptidase M Inhibitors)

  • 정명철;전효곤;이호재;이충환;고영희
    • 한국미생물·생명공학회지
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    • 제23권1호
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    • pp.47-52
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    • 1995
  • Since the original productivity of new aminopeptidase M inhibitors MR-387A and B by Streptomyces sp. SL-387 (KCTC 0102BP) was not enough for further chemical and biological evaluation, mutation of parent strain by the treatment of N-methyl-N'-nitro-N-nitrosoguanidine was performed in order to obtain a clone with greater inhibitory activity. Mutant N-3 was selected due to a 6-fold greater productivity (40 $\mu$g/ml) than that of the wild type(6.7 $\mu$g/ml). This mutant was resistant to 3,4-dehydro-DL-proline, an antimetabolite of proline, with 25 $\mu$g/ml of minimum inhibitory concentration. Furthermore, the characteristic morphological change from spiral spore chain in wild type to straight in mutant was observed. An aminopeptidase M nhibitor different from MR-387A and B was isolated from the culture broth of the mutant. This inhibitor was composed of 2 proline, 1 valine, and an unknown amino acid which is presumably 3-amino-4-phenylbutanoic acid. IC$_{50}$ value (89.1 $\MU$g/ml) of the purified inhibitor was lower than that of other inhibitors, which may be due to the absence of 2(S)-hydroxyl group within the structure of 3-amino-4-phenyl- butanoic acid.

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Label/Quencher-Free Detection of Exon Deletion Mutation in Epidermal Growth Factor Receptor Gene Using G-Quadruplex-Inducing DNA Probe

  • Kim, Hyo Ryoung;Lee, Il Joon;Kim, Dong-Eun
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.72-76
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    • 2017
  • Detection of exon 19 deletion mutation in the epidermal growth factor receptor (EGFR) gene, which results in increased and sustained phosphorylation of EGFR, is important for diagnosis and treatment guidelines in non-small-cell lung cancer. Here, we have developed a simple and convenient detection system using the interaction between G-quadruplex and fluorophore thioflavin T (ThT) for discriminating EGFR exon 19 deletion mutant DNA from wild type without a label and quencher. In the presence of exon 19 deletion mutant DNA, the probe DNAs annealed to the target sequences were transformed into G-quadruplex structure. Subsequent intercalation of ThT into the G-quadruplex resulted in a light-up fluorescence signal, which reflects the amount of mutant DNA. Due to stark differences in fluorescence intensity between mutant and wild-type DNA, we suggest that the induced G-quadruplex structure in the probe DNA can report the presence of cancer-causing deletion mutant DNAs with high sensitivity.

Pigment Reduction to Improve Photosynthetic Productivity of Rhodobacter sphaeroides

  • Kim, Nag-Jong;Lee, Jeong-Kug;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • 제14권3호
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    • pp.442-449
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    • 2004
  • Improving the light utilization efficiency of photosynthetic cells in photobioreactors (PBRs) is a major topic in algal biotechnology. Accordingly, in the current study we investigated the effect and suitability of photosynthetic pigment reduction for improving light utilization efficiency. The light-harvesting complex II (LH-II) genes of Rhodobacter sphaeroides were removed to construct a mutant strain with less pigment content. The mutant strain exhibited a slower growth rate than the wild-type under a low light intensity, while the mutant grew faster under a high light intensity. In addition, the specific absorption coefficient was lower in the mutant due to its reduced pigment content, thus it seemed that light penetrated deeper into its culture broth. However, the distance (light penetration depth) from the surface of the PBR to the compensation point did not increase, due to an increase in the compensation irradiance of the mutant strain. Experimental data showed that a reduced photosynthetic pigment content, which lessened the photoinhibition under high-intensity light, helped the volumetric productivity of photosynthetic microorganisms.