• Title/Summary/Keyword: Mutant

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

  • Gang, Hwan-Gu;Park, Hyeong-Su;Lee, Chung-Yeol;Yu, Byeong-Il;Yu, Eun-Jeong;Lee, Seon;Hwang, Seon-Deok;Gang, Hyeon-A;Lee, Sang-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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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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Site-Directed Mutagenesis of Ile91 of Restriction Endonuclease EcoRV: Dramatic Consequences on the Activity and the Properties of the Enzyme

  • Moon, Byung-Jo;Vipond, I. Barry;Halford, Stephen E.
    • BMB Reports
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    • v.29 no.1
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    • pp.17-21
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    • 1996
  • Ile91 of restriction endonuclease EcoRV, which has not been known to take part directly in catalytic activity, was substituted with Leu by site-directed mutagenesis. The Ile91Leu mutant shows over 1000-fold less activity than the wild type EcoRV under standard reaction condition. The metal ion dependency of the reaction was altered. In contrast to the wild type EcoRV, the mutant prefers $Mn^{2+}$ to $Mn^{2+}$ as the cofactor. In $Mn^{2+}$ buffer the mutant is as active as the wild type enzyme in $Mn^{2+}$ buffer. Like the wild type enzyme, the mutant shows an unspecific binding of DNA in gel shift experiments. In contrast to the wild type enzyme, the mutant did not cleave at noncognate sites of DNA under star condition.

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Generation and Evaluation of High ${\beta}$-Glucan Producing Mutant Strains of Sparassis crispa

  • Kim, Seung-Rak;Kang, Hyeon-Woo;Ro, Hyeon-Su
    • Mycobiology
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    • v.41 no.3
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    • pp.159-163
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    • 2013
  • A chemical mutagenesis technique was employed for development of mutant strains of Sparassis crispa targeting the shortened cultivation time and the high ${\beta}$-glucan content. The homogenized mycelial fragments of S. crispa IUM4010 strain were treated with 0.2 vol% methyl methanesulfonate, an alkylating agent, yielding 199 mutant strains. Subsequent screening in terms of growth and ${\beta}$-glucan content yielded two mutant strains, B4 and S7. Both mutants exhibited a significant increase in ${\beta}$-glucan productivity by producing 0.254 and 0.236 mg soluble ${\beta}$-glucan/mg dry cell weight for the B4 and S7 strains, respectively, whereas the wild type strain produced 0.102 mg soluble ${\beta}$-glucan/mg dry cell weight. The results demonstrate the usefulness of chemical mutagenesis for generation of mutant mushroom strains.

Improvement of Glyphosate Resistance through Concurrent Mutations in Three Amino Acids of the Pantoea sp. 5-Enolpyruvylshikimate-3-Phosphate Synthase

  • Liu, Feng;Cao, Yueping
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1384-1390
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    • 2018
  • Glyphosate inhibits the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in the shikimate pathway. A mutant of EPSPS from Pantoea sp. was identified using site-directed mutagenesis. The mutant showed significantly improved glyphosate resistance. The mutant had mutations in three amino acids: Gly97 to Ala, Thr 98 to Ile, and Pro 102 to Ser. These mutation sites in Escherichia coli have been studied as significant active sites of glyphosate resistance. However, in our research, they were found to jointly contribute to the improvement of glyphosate tolerance. In addition, the level of glyphosate tolerance in transgenic Arabidopsis confirmed the potentiality of the mutant in breeding glyphosate-resistant plants.

Production of Bluish Purple Pigment from Streptomyces californicus KS-89 (Streptomyces californicus KS-89 변이주에 의한 청자색소의 생산)

  • 이병호;이상훈
    • KSBB Journal
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    • v.9 no.2
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    • pp.147-156
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    • 1994
  • A study was carried out for production of a pigment : bluish purple, using a mutant Streptomyces californicus KS-89-7. The mutant was induced from Streptomyces californicus KS-89 with N-methyl-N-nitro-N-nitrosoquanidin(MNNG). It was immobilized on an inert substance made of colloidal sillica and 3.5% sodium alginate with 1 to 10 ratio. The diameter of inert bead was 2mm, and number of immobilized mutant spore was approximately $1.0{\times}10^7$/ml. It was packed in a column reactor and fermentation was conducted with a substrate made of soluble starch 1%, glycerol 1.0%, sodium glutamate 0.1%, sodium nitrate 0.05%, L-prolin 0.025% and with some trace elements. The aeration for production of the pigment was 2.5m1/min with semi-continuous fermentation. The pigment production reached at peak on 8 days of fermentation, and the mutant produced the pigment 1.8 times more than its parent strain with the maximum pigment production of $1.72g/\ell$. The pigment production continued for 24 hours of fermentation, and at the end of the fermentation the mutant produced the pigment $1.52g/\ell$.

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Ultrastructure of Ocellus in Drosophila melanogaster Visual Mutant rdgA (초파리 rdgA 시각돌연변이체 단안의 형태적 연구)

  • 윤춘식
    • Journal of Life Science
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    • v.9 no.3
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    • pp.308-313
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    • 1999
  • Ocellar morphological abnormality was studied in Drosophila rdgA mutant. In the mutant, ocellar photoreceptor cells were generally affected by the defection of rdgA molecules. Among organelles of photoreceptor cell, rhabdomeres were remarkably degenerated. The rdgA molecule, diacylglycerol kinase, was localized around SRC just below the rhabdomeric region. As a secondary phenomenon of photoreceptor degeneration, rER, multivesicular body and multilamella body were appeared in cytoplasm and these were known as to clean the cellular debris. These morphological abnormality was generally observed in degenerating cells. In Drosophila mutant, the degeneration of ocellar photoreceptor cell was facilitated to time. More intense morphological defection was observed in rdgA^{ks60} rather than in yw;rdgApc47.

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Increased Production of Recombinant Protein by Escherichia coli Deficient in Acetic Acid Formation

  • Koo, Tae-Young;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.789-793
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    • 1999
  • The effect of acetic acid formation deficiency on recombinant E. coli fermentation was investigated using a mutant strain deficient in acetic acid formation. A mutant strain which does not grow under anaerobic conditions was isolated. The acetic acid production in this strain was negligible in aerobic batch fermentation. The cloned-gene expression in the mutant strain was higher than the wild-type strain. Fed-batch fermentations with controlled specific growth rates were carried out in order to compare the cloned-gene expression between the wild-type and the mutant strains. The expression decreased along with the specific growth rate in both strains. The cloned-gene expression in the mutant strain was 60% higher than in the wild-type strain at the same specific growth rate.

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Characterization of F2 Progenies of Wound Minus Arabidopsis Mutant Crossed with Wild Type Plant

  • Park, Sanggyu
    • Journal of Applied Biological Chemistry
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    • v.43 no.1
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    • pp.12-17
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    • 2000
  • To understand the signal transduction pathway that leads to the activation of the wound-inducible proteinase inhibitor II (pin2) promoter. $F_2$ progenies of wound (-) mutant crossed with wild-type Arabidopsis plants were biochemically and genetically characterized. Wound (-) mutant was derived from transgenic Arabidopsis plants containing bacterial cytosine deaminase gene under the control of pin2 promoter. The cytosine deaminase assays indicated that wound (-) mutant is a dominant inhibitor of wound-inducibility as only 3 of the $20F_2$ progenies showed cytosine deaminase (CDase) activity, To construct a structural map of the wound (-) mutant chromosomal regions, cleaved, amplified polymorphic sequences (CAPS) markers that cover all Chromosomes were used. Chromosomal regions covered by three different CAPS markers could be candidates for further fine mapping of the location of the wound (-) mutation. g4026, RGA1 and ASA1 located at 84.9 on recombinant inbred (RI) map of chromosome I, at 1.75 on RI map of chromosome II, and 18.35 on RI map of chromosome V, respectively.

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Cloning of Cellobiohydrolase Gene (cbhI) in Radiation Induced Mutant of Pleurotus florida

  • Lee, Young-Keun;Sathesh-Prabu, Chandran;Kim, Min Kyung
    • Journal of Radiation Industry
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    • v.8 no.2
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    • pp.65-69
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    • 2014
  • The cellobiohydrolase gene (cbhI), a key component of a cellulolytic system, of a mutant PfCM4 (Pleurotus florida), developed through gamma ray radiation mutagenesis, was isolated and cloned. The deduced amino acid sequence was closely related to the glycoside hydrolase family 7 (GH7). The molecular weight of the deduced amino acid sequence of cbhI gene was found to be 22.4 kDa. Though the percent identity was found to be much less (35.61%) between the wild type and mutant, the cellulolytic activity of PfCM4 was 17.24% higher than that of the wild type. This shows that the catalytic domain of the cbhI gene was conserved in the mutant PfCM4.