• Title/Summary/Keyword: Wild strain

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An analysis of the arm-type site binding domain of bacteriophage .lambda. integrase

  • Cho, Eun-Hee
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.165-170
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    • 1995
  • The 356 amino acid long lambda integrase protein of bacteriophage .lambda. constains two autonomous DNA binding domains with distinct sequence specificities. The amino terminal domain of integrase is implicated to bind to the arm-type sequences and the carboxyl domain interacts with the coretype sequencess. As a first step to understand the molecular mechanism of the integrase-DNA interaction at the arm-type site, the int(am)94 gene carrying an amber mutation at the 94th codon of the int was cloned under the control of the P$\_$tac/ promoter and the lacI$\_$q/ gene. The Int(am)94 mutant protein of amino terminal 93 amino acid residues can be produced at high level from a suppressor free strain harboring the plasmid pInt(am)94. The arm-type binding activity of Int(am)94 were measured in vivo and in vitro. A comparison of the arm-type binding properties of the wild-type integrase and the truncated Int(am)94 mutant indicated that the truncated fragment containing 93 amino acid residues carry all the determinants for DNA binding at the arm-type sites.

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Transformation of the Edible Basidiomycete, Pleurotus ostreatus to Phleomycin Resistance

  • Kim, Beom-Gi;Joh, Jung-Ho;Yoo, Young-Bok;Magae, Yumi
    • Mycobiology
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    • v.31 no.1
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    • pp.42-45
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    • 2003
  • For transformation of Pleurotus ostreatus, two novel vectors, pPhKM1 and pPhKM2, were constructed, using the regulatory sequences of the P. sajor-caju $\beta$-tubulin gene(TUB1) and the ble gene encoding phleomycin binding protein. pPhKM1 contains ble fused to the TUB1 promoter and the Schizophyllum commune GPD terminator. pPhKM2 contains ble fused to the promoter and terminator regions of P. sajor-caju TUB1. To confirm phleomycin-resistance activity, each vector was cotrans-formed with pTRura3-2 into the P. ostreatus homokaryotic $ura^-$ strain. The transforming DNA was stably integrated into the genomic DNA. Subsequently, phleomycin resistance was conferred on wild-type dikaryotic P. ostreatus by transformation with pPhKM1 or pPhKM2. This transformation system generated stable phleomycin-resistant transformants.

Improvement of Carbapenem Antibiotics Productivity in S. cattleya by Transformation (형질전환에 의한 S. cattleya의 카바페넴 항생제 생산성 향상)

  • Park, Ji-Sun;Lee, Kang-Man
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.212-217
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    • 1996
  • Streptomyces cattleya is a producer of carbapenem antibiotics, thienamycin and N-acetylthienamycin, which have potent and broad-spectrum antibacterial activities. We stud ied on strain improvement for antibiotic productivity of S. cattleya by transformation technique which employed S.cattleya protoplasts and chromosomal DNAs of glutamic acid producers: Corynebacterium glutamicum and Arthrobacter simplex. 150 Transformant strains were cultured and bioassayed using Bacillus subtilis and Staphylococcus aureus as test organisms. 8.7% of transformants tested showed 1.4~2.6 fold higher productivities than wild type which produced $1.61{\pm}0.67{\mu}g/ml$. The best transformant produced $8.36{\pm}2.84{\mu}g/ml$ carbapenems.

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Immunological Variations of Flagella Antigens in Bacillus thuringiensis serovar kurstaki Temperature-sensitive Mutants (Bacillus thuringiensis serovar kurstaki 돌연변이 균주의 편모항원의 변이성)

  • 이형환;이영주;이희무
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.81-83
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    • 1989
  • The flagella antigenic variation of nine Bacillus thuringiensis serovar kurstaki temperature-sensitive mutants grown at the permissive temperature (3$0^{\circ}C$) was detected by a serological agglutination between H-antigen and antiserum. The flagella antigens were injected to rabbits to prepared their antisera, and then their homologous and heterologous titers of the antisera were measured. The homologous titers were ranged from 1:6,400 to 1:12,800, but the heterologous titers were very low. The H-antigen of the wild type strain was not agglutinated to 4 heterologous antisera, ts-U23 not to 7, ts-U3l not 5, ts-U32 not to 4, ts-U33 not to 7, ts-U7l not to 4, ts-U73 not to 6, ts-U74 not to 6, ts-U91 not to 4 and ts-U603 not to 4 antisera.

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Overproduction of Cellulose in Acetobacter xylinum KCCM 10100 Defective in GDP-Mannosyltransferase

  • Park Sang-Tae;Kim Eung-Bin;Kim Young-Min
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.961-964
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    • 2006
  • GDP-mannosyltransferase (GMT) is an enzyme responsible for the addition of a mannose to glucose ($\alpha$[1$\rightarrow$3]) during biosynthesis of the water-soluble branched polysaccharide acetan in Acefobacter species. In an effort to obtain a cellulose-overproducing bacterium, a mutant defective in GMT of Acetobacter xylinum KCCM 10100 was constructed by single crossover homologous recombination using part of the aceA gene encoding GMT amplified by polymerase chain reaction. The GMT-disrupted mutant produced 23% more cellulose, but 16% less water-soluble polysaccharide than those of the wild-type strain. Analysis of the sugar composition by gel permeation chromatography revealed that water-soluble polysaccharides produced by the GMT-defective mutant contained no mannose molecule.

Characteristics of a Mutant of Trehalose-producing Micrococcus luteus and Optimization of Production Conditions (Trehalose를 생산하는 Micrococcus luteus 변이주의 특성 및 생산배지의 최적화)

  • 송희상;황기철;방원기
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.399-403
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    • 1999
  • For the production of trehalose, microorganisms capable of producing trehalose extracellularly were screened from the stock cultures in our laboratory. among them, Micrococcus luteus IFO 12708 showed the highest productivity of trehalose. For the increase of productivity, the mutant strai Hs-208 having higher trehalose production was selected with NTG(N-methyl-N'-nitrosoguanidine) mutagenesis, which led to the decrease of the specific activity of trehalose phosphorylase(3.2-fold) as compared to the wild strain. The optimum condition for the trehalose production was established as follows: 20g/l of glucose and 6g/l of tryptone were used as a sole carbon source and nitrogen source, respectively, and cultivations were carried out at 3$0^{\circ}C$ and pH 6.0. After 20hrs cultivation, addition of 20unit/ml penicillin G led to the higher conversion yield of trehalose. Under the optimum condition, 6.547g/l trehalose was produced with conversion yield of 32.7%.

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Studies on the Susceptibility of Insecticides and Killing Effect by Thermal Fogging Against Culex pipiens pallens of Ansan Strain (안산시 서식 빨간집모기(Culex pipiens pallens)에 대한 살충제 감수성 및 가열연막 효과에 관한 연구)

  • Choi Han-Young
    • Journal of environmental and Sanitary engineering
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    • v.20 no.3 s.57
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    • pp.44-50
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    • 2005
  • Studies on the insecticides susceptibility of adults females Culex pipiens pallens were carried out in 2003. The pupae were emerged originated wild-caught larvae in Ansan city, Korea. The test methods employed, using 7 organophosphorous insecticides, four synthetic pyrethroides, and fipronil penyrazole were evaluated. The following results were obtained 1. Fipronil has showed the most strong value in $LD_{50}\;0.00075{\mu}g/female$, out of 12 kind of insecticides, and followed by deltamethrin 0.0071, $\delta-cyhalothrin\;0.008$, profenofos 0.0082 and $\beta-cyfluthrin$ 0.0088, respectively 2. The killing effect of lambdacyhalothrin and profenophos against adult females Culex pipiens pallens was examined using thermal fogging. The mortality rate were lambdacyhalothrin $41.1\%$ and profenophos $50.7\%$, respectively. The killing effect of thermal fogging was highly effectiveness to distance 6m from nozzle

Genetic Analysis on Bioconversion of Aniline to Acetaminophen in Streptomyces fradiae

  • Jin, Hyung-Jong;Park, Ae-Kyung;Lee, Sang-Sup
    • Archives of Pharmacal Research
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    • v.15 no.1
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    • pp.35-40
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    • 1992
  • S. fradiae showed the highest acetanilide p-hydroxylation activity in the tested strains. And S. fradiae was well characterized genetically, especially with respect to tylosin production. Two mutants, which lost hydroxylation, were isolated in 140 regenerated colonies from protoplasts. In restriction enzyme digesion of total DNAs, isolation of giant linear plasmid DNA and determination of antibiotic resistances to chloramphenicol, tylosin, hygromycin B and mitomycin C, any differences among mutants and a wild type strain were not detected. These facts suggest that lesion on 6, 000 Kb chromosomal DNA was responsible for the lack of p-hydroxylation activity induced by protoplast formation and regeneration.

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Roles of the meta- and the ortho-Cleavage Pathways for the Efficient Utilization of Aromatic Hydrocarbons by Sphingomonas yanoikuyae Bl

  • 송정민;김영민;Gerben J. Zylstra;김응빈
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.245-245
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    • 2002
  • Catabolic pathways for the degradation of various aromatics by Sphingomonas yanoikuyae Bl are intertwined, joining at the level of substituted benzoates, which are further degraded vita ring cleavage reactions. The mutant strain EK497, which was constructed by deleting a large DNA region containing most of the genes for biphenyl, naphthalene, m-xylene, and m-toluate degradation, was unable to grow on all of the aromatics tested except for benzoate as the sole source of carbon and energy.S. yanoikuyae EK497 was found to possess only catechol ortho-ring cleavage activity due to deletion of the genes for the meta-cleavage pathway. Wild-type S. yanoikuyae Bl grown on benzoate has both catechol orthoand meta-cleavage activity. However, m-xylene and m-toluate, which are metabolized through methylbenzoate, and biphenyl, which is metabolized through benzoate, induce only the meta-cleavage pathway, suggesting the presence of a substrate-dependent induction mechanism.

Colistin 생산균주의 균주개량 및 productivity 증대를 위한 발효최적화

  • Ye, Byeong-Dae;Hwang, Yong-Bae;Kim, Yeong-Hui;Kim, Dong-Geon;Yang, Ho-Seok
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.224-227
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    • 2002
  • Colistin produced from Penibacillius polymyxa was widely used as an antibiotic active against gram-negative bacteria and as feed additive. This research studied on increment of colistin productivity by mutation of P. polymyxa. As a result, several mutants were obtained from the strain by UV radiation and NTG treatment. They produced approximately 8.5${\sim}$9.0 g/L of colistin in flask and jar culture. Colistin productivity of the mutant, named Penibacillius polymyxa CBY, showed 100 times than that of wild type. When Penibacillius polymyxa CBY fermented in the optimal medium, it produced up to 18 g/L of colistin in jar fermentation.

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