• Title/Summary/Keyword: Wild strain

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R-plasmids in staphylococcus aureus (Staphylococcus aureus의 항생제 내성 plasmid에 관한 연구)

  • 변우현;김영선;조은희;권동현;이호주;홍순주
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.282-290
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    • 1985
  • Small size antibiotic resistance plasmids having molecular weights less than 10 Mdal were isolated and characterized from ten clinically isolated multiple resistant Staphylococcus aureus. Agarose gel electrophoresis profiles and antibiotic resistance patterns divided these strains into four groups. Strain 2-23-6, the representative strain of a group of five strains conferred two plasmids of molecular weights $1.6{\times}10^6\;dal\;and\;2.0{\times}10^6$ dal. The small plasmid (pSBK 112) specified macrolides, lincosamides and streptogramin type B (MLS) resistance gene which are expressed constitutively. Lage plasmid (pSBK 125) specified chloramphenicol resistance gene which is inducible. Strain 10-5 conferred a $3.0{\times}10^6$ dal plasmid (pSBK 141) which carry an inducible ampicillin resistance gene and strain P-H-2 conferred and $1.6{\times}10^6$ dal plasmid (pSBK 190) which carry a constitutive MLS resistance gene. Strain D-H-1 conferred four plasmids of molecular weights $0.8{\times}10^6$ dal (pSBK 201), $1.6{\times}10^6$ dal (pSBK 202), $2.5{\times}10^6$ dal (pSBK 203), and $1.2{\times}10^7$ dal (pDBK 204), respectively. Among those four plasmids, only pSBK 203 specified chloramphenicol resistance gene. Curing of constitutive MLS resistance using acriding orange or ethidium bromide in 2-23-6 and P-H-2 strains produced 'inducible' MLS resistance strains which are less resistant to MLS than the wild type strains, suggesting that there are two resistance genes in both strains; one is constitutive and the other is inducible.

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Detection of Zymogenic ChsC Activity in Vegetative Hyphae of Aspergillus nidulans. (Aspergillus nidulans 영양균사에서 효소전구체형 ChsC 활성의 검출)

  • 박범찬;박윤희;박희문
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.178-182
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    • 2004
  • In the vegetative hyphae of Aspergillus nidulans, a zymogenic form of the class I chitin synthase activity was successfully measured by the assay condition for Saccharomyces cerevisiae class I chitin synthase, Chsl. The class I chitin synthase activity of the A. nidulans chsC wild type strain was increased about six-fold by trypsin-pretreatment, but that of the chsC disruption strain revealed no increase. Interestingly enough, level of the class I chitin synthase activity of the chsC disruption strain was almost the same as that of the chsC wild type without trypsin-pretreatment. These results indicated that the A. nidulans ChsC activity could be measured by account-ing the class I chitin synthase activity without the trypsin-pretreatment as an internal control. Consistence to the expression pattern of the chsC revealed by northern blot analysis, the activity of ChsC was increased upon reaching the culture time for acquiring developmental competence. Our results shown here also supported the previous report suggesting the possible involvement of ChsC in vegetative hyphal growth of A. nidulans.

Genetic Engineering of Biological Nitrogen Fixation and its Application to Agronomy - Selection of Rhizobium japonicum Mutants having Greater Symbiotic Nitrogen Fixing Activity with Soybean - (질소고정(窒素固定)의 유전공학(遺傳工學的) 연구(硏究) 및 농업(農業)에의 응용방안(應用方案) - 대두(大豆)에 효율적인 공생질소고정(共生窒素固定)을 할 수 있는 Rhizobium japonicum mutant의 선별 -)

  • Cho, Moo Je;Yang, Min Suk;Yun, Han Dae;Choe, Zhin Ryong;Choe, Yong Lark;Kang, Kyu Young
    • Microbiology and Biotechnology Letters
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    • v.13 no.1
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    • pp.79-85
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    • 1985
  • Rhizobium japonicum wild type strains isolated from local soybean variety Jangback root nodules with higher nitrogenase activity than R. japonicum 3I1110 or 61A76 was mutangenized by N-methyl-N'-nitro-N-nitrosoguanidine and UV-irradiation, and screened by effectiveness assay with soybean. One mutant strain JB65 nodulated the roots earlier than the wild type and also expressed higher acetylene-reducing activity in the presence and absence of fixed nitrogen. The selected mutant was compared with SM35 strain and showed greater nodulation and symbiotic nitrogen fixing activity with local soybean variety Jangback than SM35 strain.

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Control of Genes in TCA Cycle by fsrA Small RNA in Bacillus subtilis (바실러스 서브틸리스의 fsrA small RNA에 의한 TCA 회로의 유전자 조절)

  • Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.19 no.1
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    • pp.57-64
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    • 2008
  • The fsrA gene in Bacillus subtilis has an analogous role of ryhB in E. coli and is controlled under fur, the iron regulator gene. At high concentration of iron the transcription of ryhB is repressed by fur and ryhB is transcribed under low concentration of iron. To spare iron produced ryhB small RNA represses the expression of sdhCDAB (succinate dehydrogenase). This study shows the growth rate of Bacillus subtilis strain of fur and fur/fsrA deletion mutants using organic acids of TCA cycle as carbon source. Mutant strain of fur does not grow well with succinate carbon source, but further deletion of fsrA regain to the growth of wild type strain. Also, nearly same results were observed with citrate and fumarate. These results are consistent to those of E. coli system. But fur and fur/fsrA deletion mutants grow well as much as the growth of wild type with malate carbon source. These results showed that upstream genes of succinate of TCA cycle are repressed by fsrA, but downstream of succinate are not repressed by fsrA.

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Putative response regulator two-component gene, CaSKN7, regulate differentiation and virulence in Candida albicans

  • Lee, Jung-Shin;Minyoung Lim;Yim, Hyung-Soon;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.50-50
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    • 2003
  • We have identified and analysed a putative response regulator two-component gene (CaSKN7) from Candida albicans and its encoding protein (CaSkn7). CaSKN7 has an open reading frame of 1677bp. CaSKN7 encodes a 559 amino acid protein (CaSkn7) with an estimated molecular mass of 61.1 kDa. CaSKN7 is a homologue of a Saccharomyces cerevisiae SKN7 that is the regulator involved in the oxidative stress response. To study the role of CaSKN7, we constructed a CAI4-derived mutant strain carrying a homozygous deletion of the CaSKN7 gene. In the caskn7 disruptant cells, the formation of germ tube require shorter time than that in the congenic wild-type strain but the growth of mycelium delayed in liquid media. In contrast, the caskn7 disruptant cells attenuate the differentiation in solid media and the virulence in mouse model system. Expression level of hypha-specific and virulence genes - HYR1, ECE1, HWP1, and ALS1 - in the caskn7 disruptant cells increased as compared with that in the congenic wild-type strain in 10% serum YPD. Skn7 in 5. cerevisiae was found to bind the HSE element from the SSA promoter, Also, CaSkn7 contains heat shock factor DNA-binding domain and the promoters of these genes have HSE-like sties. Therefore these results show that CaSKN7 regulate the differentiation and virulence of C. albicans.

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Deletion of cg1360 Affects ATP Synthase Function and Enhances Production of L-Valine in Corynebacterium glutamicum

  • Wang, Xiaochen;Yang, Hongyu;Zhou, Wei;Liu, Jun;Xu, Ning
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1288-1298
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    • 2019
  • Bacterial ATP synthases drive ATP synthesis by a rotary mechanism, and play a vital role in physiology and cell metabolism. Corynebacterium glutamicum is well known as an industrial workhorse for amino acid production, and its ATP synthase operon contains eight structural genes and two adjacent genes, cg1360 and cg1361. So far, the physiological functions of Cg1360 (GenBank CAF19908) and Cg1361 (GenBank CAF19909) remain unclear. Here, we showed that Cg1360 was a hydrophobic protein with four transmembrane helices (TMHs), while no TMH was found in Cg1361. Deletion of cg1360, but not cg1361, led to significantly reduced cell growth using glucose and acetic acid as carbon sources, reduced F1 portions in the membrane, reduced ATP-driven proton-pumping activity and ATPase activity, suggesting that Cg1360 plays an important role in ATP synthase function. The intracellular ATP concentration in the ${\Delta}cg1360$ mutant was decreased to 72% of the wild type, while the NADH and NADPH levels in the ${\Delta}cg1360$ mutant were increased by 29% and 26%, respectively. However, the ${\Delta}cg1361$ mutant exhibited comparable intracellular ATP, NADH and NADPH levels with the wild-type strain. Moreover, the effect of cg1360 deletion on L-valine production was examined in the L-valine-producing V-10 strain. The final production of L-valine in the $V-10-{\Delta}cg1360$ mutant reached $9.2{\pm}0.3g/l$ in shake flasks, which was 14% higher than that of the V-10 strain. Thus, Cg1360 can be used as an effective engineering target by altering energy metabolism for the enhancement of amino acid production in C. glutamicum.

Hydrogen Cyanide Produced by Pseudomonas chlororaphis O6 Exhibits Nematicidal Activity against Meloidogyne hapla

  • Kang, Beom Ryong;Anderson, Anne J.;Kim, Young Cheol
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.35-43
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    • 2018
  • Root-knot nematodes (Meloidogyne spp.) are parasites that attack many field crops and orchard trees, and affect both the quantity and quality of the products. A root-colonizing bacterium, Pseudomonas chlororaphis O6, possesses beneficial traits including strong nematicidal activity. To determine the molecular mechanisms involved in the nematicidal activity of P. chlororaphis O6, we constructed two mutants; one lacking hydrogen cyanide production, and a second lacking an insecticidal toxin, FitD. Root drenching with wild-type P. chlororaphis O6 cells caused juvenile mortality in vitro and in planta. Efficacy was not altered in the fitD mutant compared to the wild-type but was reduced in both bioassays for the mutant lacking hydrogen cyanide production. The reduced number of galls on tomato plants caused by the wild-type strain was comparable to that of a standard chemical nematicide. These findings suggest that hydrogen cyanide-producing root colonizers, such as P. chlororaphis O6, could be formulated as "green" nematicides that are compatible with many crops and offer agricultural sustainability.

ANTIBIOTIC SUSCEPTIBILITY IN MUTANS STREPTOCOCCI AND STREPTOCOCCUS ANGINOSUS ISOLATED FROM DENTAL PLAQUE (치면세균막에서 분리한 뮤탄스 연쇄상구균 및 Streptococcus anginosus의 수종 항생제에 대한 감수성 조사)

  • Kook, Joong-Ki;Lim, Sang-Soo;Yoo, So-Young;Hwang, Ho-Keel
    • Restorative Dentistry and Endodontics
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    • v.29 no.5
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    • pp.462-469
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    • 2004
  • The aim of this study was to investigate the susceptibility of mutans streptococci (S. mutans and S. sobrinus) and Streptococcus anginosus, for seven antibiotics, penicillin G, amoxicillin, ciprofloxacin, cefuroxime, erythromycin, bacitracin, and vancomycin. The minimum inhibitory concentration (MIC) of seven antibiotics against 3 species (type strains) of mutans streptococci and S. anginosus, 10 strains (wild type) of S. mutans, 7 strains (wild type) of S. sobrinus, and 11 strains (wild type) of S. anginosus, were measured by broth dilution method. All of the type strains of mutans streptococci and S. anginosus had the same susceptibility for penicillin G, amoxicillin, cefuroxime and bacitracin. Type strain of S. anginosus was sensitive in ciprofloxacin, but those of mutans streptococci were not. All of the clinical isolates of mutans streptococci and S. anginosus had the same susceptibility for the seven antibiotics. Our data reveal that mutans streptococci and S. anginosus have similar antibiotic-resistant character. In addition. these results may offer the basic data to verify the antibiotic-resistant mechanism of mutans streptococci and S. anginosus.

Characterization of Complemented Mutants in Pseudomonas fluorescens and Cloning of the DNA Region Related in Antibiotic Biosynthesis (길항세균 Pseudomonas fluorescens의 Complemented Mutant에 대한 특성조사에 및 길항물질 유전자 Cloning)

  • Kim, Young;Cho, Yong-Sup
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.151-156
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    • 1994
  • Pseudomonas fluorescens produces the antibiotic, 2,4-diacetylphloroglucinol (Phl), which promotes plant growth by inhibiting bacteria and fungi. Cosmids (genomic library) were mobilized into Phl-nonproducing mutants through the triparental matings with pRK2013 as the helper plasmid at the frequency of 8.37$\times$10-4. Complemented mutants that showed antibiotic activity were selected among about 2,000 transconjugants. The complemented mutants were confirmed by acquired drug resistances (kanamycin and tetracycline). The antibiotic substances of wild type and complemented mutants showed the most excellent anti-bacterial activity. Inhibitory effects of complemented P. fluorescens against phytopathogenic fungi were equal to the parental strain. Complemented mutant and wild type of P. fluorescens were causal microbes of fungal morphological abnormalities. Complemented mutants in potato dextrose agar supplemented with bromothymol blue also showed restoration of glucose utilization as wild type. Plasmids of complemented mutants were isolated from transconjugant sand transformed into competent cells of E. coli DH5$\alpha$. The plamid DNA was reisolated from transformed E. coli DH5$\alpha$.

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Potential Yeast from Indonesian Wild Forest Honey Showing Ability to Produce Lipase for Lipid Transesterification

  • Palilu, Prayolga Toban;Kasiamdari, Rina Sri;Ilmi, Miftahul
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.555-564
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    • 2019
  • Biodiesel is produced through the transesterification process in the presence of alcohol and a catalyst that catalyzes the conversion of triglycerides to esters and glycerol compounds. A more optimal product conversion can be achieved using enzymes, such as lipase. Lipase is reported to be produced in osmophilic yeasts due to the low water content in their natural habitats. Wild forest honey is one of the osmophilic natural habitats in Indonesia. However, lipase-producing yeast has not been reported in the Indonesian honey. In this study, we screened the lipase-producing yeasts isolated from wild forest honey collected from Central Sulawesi. The production profile and activity of lipase were determined at different pH values and temperatures. One promising yeast was isolated from the honey, which was identified as Zygosaccharomyces mellis SG 1.2 based on ITS sequence. The maximum lipase production (24.56 ± 1.30 U/mg biomass) was achieved by culturing the strain in a medium containing 2% olive oil as a carbon source at pH 7 and 30℃ for 40 h. The optimum pH and temperature for lipase activity were 6 and 55℃, respectively. The enzyme maintained 80% of its activity upon incubation at 25℃ for 4 h. However, the enzyme activity decreased by more than 50% upon incubation at 35 and 40℃ for 2 h. This is the first study to report the lipase producing capability of Z. mellis. Further studies are needed to optimize the enzyme production.