• Title/Summary/Keyword: Nucleic acids

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Mesenchymal Stem Cell-derived Exosomes: Applications in Cell-free Therapy (중간엽줄기세포유래 엑소좀: 비세포치료제로서의 활용)

  • Heo, June Seok;Kim, Jinkwan
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.391-398
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    • 2018
  • Mesenchymal stem cells (MSCs) are an attractive resource for refractory patients because of their anti-inflammatory/immunomodulatory capability and multi-lineage differentiation potential. The transplantation of MSCs has led to positive results in preclinical and clinical application to various diseases, including autoimmune disease, cardiovascular disease, cancer, liver cirrhosis, and ischemic stroke. On the other hand, studies have shown that paracrine factors, not direct cell replacement for damaged cells or tissue, are the main contributors in MSC-based therapy. More recently, evidence has indicated that MSC-derived exosomes play crucial roles in regulating the paracrine factors that can mediate tissue regeneration via transferring nucleic acids, proteins, and lipids to the local microenvironment and cell-to-cell communication. The use of these exosomes is likely to be beneficial for the therapeutic application of MSCs because their use can avoid harmful effects, such as tumor formation involved in cell transplantation. Therefore, therapeutic applications using MSC-derived exosomes might be safe and efficient strategies for regenerative medicine and tissue engineering. This review summarizes the recent advances and provides a comprehensive understanding of the role of MSC-derived exosomes as a therapeutic agent.

Characterization of Three Korean Isolates of Malva Vein Clearing Virus from Curled Mallow (Malva verticillata) (아욱에서 분리한 Malva Vein Clearing Virus 분리주의 특성)

  • Kwak, Hae-Ryun;Kim, Ji-Gwang;Kim, Jeong-Eun;Choi, Hyeon-Yong;Choi, Hong-Soo;Kim, Mikyeong
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.283-288
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    • 2020
  • In September 2017, vein clearing and yellowing symptoms resembling those caused by viruses were observed on leaves of Malva verticillata in Chungnam, Korea. Nucleic acids were extracted from leaves of five symptomatic plants and tested by reverse transcription polymerase chain reaction using four virus specific primer pairs including malva vein clearing virus (MVCV). Amplicons of the expected size (600 bp) were obtained from total RNA of all samples using the MVCV-specific primers. To confirm the presence of MVCV in symptomatic plants, the DNA fragments from three samples were purified, and directly sequenced. BLAST analysis revealed that it shared the highest nucleotide identity (99%) with a MVCV isolate from tomato (Mexico). The virus isolates obtained from the third re-inoculated Chenopodium was designated as Cm1-5. Tissue from Cm1, Cm3, and Cm5 isolates was mechanically sap inoculated into 23 indicator plants. Cm3 isolate induced chlorotic local and mosaic symptoms in Althaea rosea. Phylogenetic analysis based on coat protein gene of 19 MVCV isolates from 6 different countries and plant species, did not correlated with either the geographical origin of the isolates, or pathogenicity. To our knowledge, this study first reports the natural occurrence of MVCV on M. verticillata in Korea and characterization of three Korean isolates of MVCV.

Preparation and Characterization of Lipid Nanoparticles Containing Fat-Soluble Vitamin C Derivatives and Gallic Acid (지용성 비타민 C 유도체 및 갈릭산을 함유한 지질나노입자 제조 및 특성)

  • Ji Soo Ryu;Ja In Kim;Jae Yong Seo;Young-Ah Park;Yu-Jin Kang;Ji Soo Han;Jin Woong Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.2
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    • pp.103-110
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    • 2024
  • Lipid nanoparticles (LNPs) are a stable and an effective system that protects cell-impermeable biologically active compounds such as nucleic acids, proteins, and peptides against degradation caused by subtle environmental changes. This study focuses on developing LNPs encapsulating gallic acid (GA), an antioxidant, to effectively prolong the half-life of tetrahexyldecyl ascorbate (THDC), a oil-soluble vitamin C derivative. These LNPs were synthesized in small, uniform sizes at room temperature and pressure conditions using a microfluidics chip. Compared to liposomes manufactured under high pressure and high temperature conditions through conventional microfluidizers, LNPs manufactured through microfluidics chips had excellent dispersion and temperature stability, and improved skin absorption as well as improved oxidative stability of fat-soluble vitamin C derivatives. Future studies will focus on ex vivo and in vivo evaluations to study skin improvement to further validate these results.

Major Components of Teas Manufactured with Leaf and Flower of Korean Native Camellia japonica L. (국내 자생 동백나무의 잎과 꽃으로 만든 엽차와 화차의 주요성분)

  • Cha, Young-Ju;Lee, Jang-Won;Kim, Ju-Hee;Park, Min-Hee;Lee, Sook-Young
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.3
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    • pp.183-190
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    • 2004
  • The major compositions of leaf tea and flower tea were investigated to develope as a new functional tea using Korean native Camellia japonica L. Most of leaf teas, except flower tea, were considered as good materials with basic conditions for tea manufacture because water content was below 6%. Crude protein was the greatest component in roasted young leaf tea (RYLT), crude fats in roasted mature leaf tea (RMLT) and ashes in fermented young leaf tea (FYLT). Caffein were present as the highest amount (5.18%) in steamed mature leaf tea (SMLT), showing less amount than green tea. Catechin were contained as the highest amount in all kinds of teas, especially FYLT was the highest (9.57%). Tannin, which highly related with tea quality including astringent taste, color and perfume, were present as the highest amount in FYLT. Vitamin C was highly detected in the tea from flowers (22.7 mg/l00 g) rather than in the tea from leaves. The content of theanine were found in flower tea by 1,074 mg/l00 g, and had about twofold of FYLT and RYLT. Among free amino acids, glutamic acid and aspartic acid were higher detected in SMLT and RMLT while asparagine was present as higher amounts in RYLT and FYLT, expecting these components can improve tea taste. Nucleic acids and their derivatives including GMP, hypoxanthine and AMP were detected as the higher amounts by 7.86, 8.57, and $12.67\;{\mu}mol/g$, respectively, however IMP content was even reduced by all manufacturing processes. In all kinds of tea, sugars such as glucose, fructose, sucrose and maltose were detected, specially glucose and fructose were found as highest amount in RFT by 65.5 and 59.6 nmol/0.1 mg, respectively.

Studies on Degradation of Nucleic acid and Related Compounds by Microbial Enzymes (미생물 효소에 의한 핵산 및 그의 관련물질의 분해에 관한 연구)

  • Kim, Sang-Soon
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.111-129
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    • 1970
  • As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.

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Discrimination of African Yams Containing High Functional Compounds Using FT-IR Fingerprinting Combined by Multivariate Analysis and Quantitative Prediction of Functional Compounds by PLS Regression Modeling (FT-IR 스펙트럼 데이터의 다변량 통계분석을 이용한 고기능성 아프리칸 얌 식별 및 기능성 성분 함량 예측 모델링)

  • Song, Seung Yeob;Jie, Eun Yee;Ahn, Myung Suk;Kim, Dong Jin;Kim, In Jung;Kim, Suk Weon
    • Horticultural Science & Technology
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    • v.32 no.1
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    • pp.105-114
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    • 2014
  • We established a high throughput screening system of African yam tuber lines which contain high contents of total carotenoids, flavonoids, and phenolic compounds using ultraviolet-visible (UV-VIS) spectroscopy and Fourier transform infrared (FT-IR) spectroscopy in combination with multivariate analysis. The total carotenoids contents from 62 African yam tubers varied from 0.01 to $0.91{\mu}g{\cdot}g^{-1}$ dry weight (wt). The total flavonoids and phenolic compounds also varied from 12.9 to $229{\mu}g{\cdot}g^{-1}$ and from 0.29 to $5.2mg{\cdot}g^{-1}$dry wt. FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and $1,100-950cm^{-1}$, respectively. These spectral regions were reflecting the quantitative and qualitative variations of amide I, II from amino acids and proteins ($1,700-1,500cm^{-1}$), phosphodiester groups from nucleic acid and phospholipid ($1,500-1,300cm^{-1}$) and carbohydrate compounds ($1,100-950cm^{-1}$). Principal component analysis (PCA) and subsequent partial least square-discriminant analysis (PLS-DA) were able to discriminate the 62 African yam tuber lines into three separate clusters corresponding to their taxonomic relationship. The quantitative prediction modeling of total carotenoids, flavonoids, and phenolic compounds from African yam tuber lines were established using partial least square regression algorithm from FT-IR spectra. The regression coefficients ($R^2$) between predicted values and estimated values of total carotenoids, flavonoids and phenolic compounds were 0.83, 0.86, and 0.72, respectively. These results showed that quantitative predictions of total carotenoids, flavonoids, and phenolic compounds were possible from FT-IR spectra of African yam tuber lines with higher accuracy. Therefore we suggested that quantitative prediction system established in this study could be applied as a rapid selection tool for high yielding African yam lines.

Autometallography for Zinc Detection in the Central Nervous System (중추신경계통내 분포하는 Zinc의 조직화학적 동정)

  • Jo, Seung-Mook;Gorm, Danscher;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
    • Applied Microscopy
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    • v.30 no.4
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    • pp.347-355
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    • 2000
  • Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to some metalloproteins and nucleic acids, loosely bound to some metallothioneins or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors and transcription factors, thus rolls need accurate homeostasis of zinc ions. Zinc is an essential catalytic or structural element of many proteins, and a signaling messenger that is released by neural activity at many central excitatory synapses. Growing evidences suggest that zinc may also be a key mediator and modulator of the neuronal death associated with transient global ischemia and sustained seizures, as well as perhaps other neurological disease stoles. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartment ('vesicular zinc') which can be evidenced using histochemical techniques. Substances giving a bright colour or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called 'direct' methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process and, finally, their demonstration after autometallographic development are essential steps for other methods, the so called 'indirect methods'. This study is a short report on the autometallograhical approaches for zinc detection in the central nervous system (CNS) by means of a modified selenium method.

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Studies on the Bacteriophages of Brevibacterium lactofermentum (L-글루타민산 생산균 Brevibacterium lactofermentum의 Bacteriophag에 관한 연구)

  • 이태우
    • Korean Journal of Microbiology
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    • v.17 no.3
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    • pp.97-130
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    • 1979
  • Many industrial processes those employ bacteria are subjected to phage infestations. In L-glutamic acid fermentions using acetic acid, the phage infestations of the organisms have been recently recognized. In efforts to elucidate the sources of phage contamination involved in the abnormal fermentation, a series of study was conducted to isolate the phages both from the contents of abnormally fermented tanks and the soil or sewage samples from the surroundings of a fermentation factory, to define major charateristics of the phage isolates, and finally to determine the correlation between the phage isolates and temperate phages originating from the miscellaneous bacterial species isolated from the soil or sewage samples. The results are summarized as follows; 1) All phages were isolated from the irregular fermentation tanks and soil or sewage samples, and they were designated as phage PR-1, PR-2, PR-3, PR-4, PR-5, PR-6, and PR-7, in the order of isolation. These PR-series phages were proved to be highly specific for the variant strains of Br. lactofermentum only, namely, phage PR-1 and PR-2 for Br. lactofermentum No. 468-5 and phage PR-3~PR-7 for Br. lactofemrentum No. 2256. By cross-neutralization test, the 7 phagescould be subdivided into 3 groups, i. e., phage PR-I and PR-2 the first, phage PR-3, PR-4, PR-5, PR-6 the second, and the phage PR-7 the third. 2) The 7 phages were virulent under the experimental conditions. They produced plaques with clear and relatively sharp margins without distinct halo. The mean sizes of plaques were 1.5mm in diameter for phage PR-1 and PR-2, and 1. Omm for phages PR-3~PR-7. Double layer technique modified by Hongo and described by Adams, was applied to assay of the PR-series phages. The factors influencing the plaques were as follows;young age cells of host bacteria cultured for 3-6 hours represented the largest number and size, optimum was pH 7.0, incubation temperature was $30^{\circ}C$, and agar concentration and amount of overlayer medium were 0.6% and 0.2ml, respectively. 3) PR-series phages were stable in 0.05M tris buffer and 0.1M ammonium acetate buffer solution. The addition of $5{\times}10^{-3}M$ magnesium ion effectively increased the stability. Thermostability experiments indicated that PR-series phages were stable at the teinperture between $50^{\circ}{\sim}55^{\circ}C$ in nutrient medium, $45^{\circ}{\sim}50^{\circ}C$ in buffer solution. However, the phages mere completely inactivated at 603C and 65$^{\circ}$C within 10 minutes. The phages were stable at the range of pH6~9 in nutrient medium and of pH 8-9 in buffer solution, respectively. Exposure of the phages to UV for 25, 60 and 100 seconds resulted in the complete loss of infectivily, respectively. 4) Electron microscopy showed that PR-series phage particles exhibited rather similar morphology, differing in the size All of PR-series phages had a multilateral head and had a simple long tiil about three to five times long as compared with head. By the size, phage PR-1 and PR-2, PR-3, PR-4, PR-5, and PR-6 and PR-7 were classified into same groups, respectively. The head and tail size of phage PR-1, PR-5, PR-5(T) and PR-7 were 85nm, 74nm and 235nm and 350mm, and 72nm and 210nm, respectively. 5) Nucleic acids of PR-series phages were double stranded DNA. The G+C contents of phage PR-1, PR-5 and PR-7 were 56.1, 52.9 and 53.7, respectively. The values of G+C contents derived from the $T_m$ were in agreement with the chemically determined values. 6) PR-series phages effectively adsorbed on their host bacteria at the rate of more than 90% during 5 min. K value for phage PR-1, PR-5 and PR-7 were calculated to be $6{\times}10^9 ml$ per minute, respectiveky. The pH of the medium did effect adsorption rate, but both temperature and age of host cells did not. Generally, optimum adsorption condition of phages seemed to be almost same as optimum growth conditions of host bacteria. 7) In one-step growth experiments, the latent periods at $30^{\circ}C$ for PR-1, and PR-7 were about 70, 50 and 55 min, respectively. The corresponding average burst size was 200, 70 and 90, respectively. Lpsis period according to the multiplicity of infection and a phage series. In case of m. o. i. 100, strain No. 2256 (PR-5) and No. 468-5(PR-1) failed to grow and turbidity decreased after 50 and 70min, respectively. 8) In the lysate of a plaque purified phage PR-5 infected bacteria, there observed 2 types ofphage particles, i. e., phage PR-5 and PR-5 (T) of similar morphology but differing at the length of phage tail, and phage tail like particles. The phage taillike particles could be divided into 4 types by the length. Induction experiments of Br. lactofermentum with UV irradiation, mitomycin C or bacitracin treatment produced neither phage PR-5 (T) or phage tail-like particles. 9) No lysis occured when the growth of 7 strains of miscellaneous bacteria, isolated from soil and sewage samples, were inoculated with either phage PR-5 (T) or phage tail-like particles the inoculation of phage PR-5 pellet resulted in the growth inhibition of the orgainsms in the spot test. The lysates obtained from 3 miscellaneous soil derived bacteria following mitomycin C treatment the growth of Br. lactofermentum, but did not lyze the bacterium.

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Effects of Vitamins and Nucleic acids on the mycelial growth and the sclerotial production of Sclerotium rolfsii (Vitamin과 핵산이 Sclerotium rolfsii의 균사생장 및 균핵형성에 미치는 영향)

  • KIM KI CHUNG
    • Korean journal of applied entomology
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    • v.12 no.2
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    • pp.71-78
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    • 1973
  • The study was performed to clear the effects of thiamine, biotin, nicotinic acid, pyridoxine, inositol, deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) on the mycelial growth and the sclerotial production of Sclerotium rolfsii Sacc. isolated from Magnolia kobus. The results are abstracted as follows: 1. Tested fungus was thiamine- deficient and required thiamine 20r/l for maximum growth of mycelia. At higher concentrations than thiamine 20r/l, however, mycelial growth was decreased with increasing the concentrations and was inhibited little less than that of thiamine-free control at 150r/l. 2. The effecfivenesses of the nitrogen sources on the mycelial growth under the thiamine presence were recognized in order of $NH_4NO_3>(NH_4)_2SO_4 >asparagine> KNO_3$, and on the sclerotial production were $KNO_3>NH_4NO_3>asparagine>(NH_4)_2SO_4$. The optimum concentrations of thiamine were about 12r/1 in $KNO_3$, about 16r/1 in asparagine on the growth of mycelia, and were about 8r/l in $KNO_3\;and\; NH_4NO_3,\; 16r/1$ in asparagine on the production of sclerotia. 3. After the organism began to grow, the pH value of cultral filtrate was rapidly dropped down to about 3.5. Hereafter it was slowly fallen down as the growth amount was increased, but was not depreciated below pH 2.2. 4. Nicotinic acid was not effective individually on the mycelial growth and the sclerotial formation of tested fungus without thiamine, but slight effect of it was recognized with thiamine 10r/l, even though maximum growth was shown at 7-10mg/1. Beyond that concentration, however, mycelial growth was rather depressed. 5. When ammonium sulphate or asparagine as the nitrogen sources was used, pyridoxine, biotin and inositol had not any effectivenesses on the mycelial growth and the sclerotial production of examined fungus. 6. In the concentrations of thiamine, biotin, pyridoxine and inositol, as long as thiamine was not added in those, their correlating effects on the growth of the organism were not observed at all. Equivalent or more effects on the mycelial growth were recognized in combinations of thiamin + pyridoxine, thiamine + inositol, thiamine + biotin + pyridoxine, and thiamine + biotin + pyridoxine + inositol compared with thiamino alone, and in combinations of thiamine + biotin and thiamine + biotin + inositol, mycelial growth was inhibited rather than that of thiamine alone. Sclerotial production of those combinations was increased more than that of thiamine alone in dry weight. 7, The little effects of DNA and RNA on the mycelial growth of the organism were recognized compared with the control(DNA-and RNA-free), and RNA was more effective than DNA. Maximum growth of mycelia was observed at RNA 2-6mg/1 and DNA 6mg/l. No effectivenesses on the sclerotial production were recognized in the RNA and DNA. 8. Mycelial growth of the organism was increased with increasing the concentrations of the RNA and the thiamine, that is, the effectiveness of RNA was revealed apparently under presence of thiamine, but was not shown in the sclerotial formation.

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Application of a New Conjugation Method to Fish Pathogenic Bacteria Containing R Plasmid for the Analysis of Drug-Resistant Status in Aquaculture (새로운 conjugation 방법을 응용한 R plasmid 함유 어병세균의 분리와 양식장 내성균의 현황 분석)

  • Yoo Min Ho;Jeong Joon Beom;Kim Eun Heui;Lee Hyoung Ho;Jeong Hun Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.2
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    • pp.115-121
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    • 2002
  • To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.