• 제목/요약/키워드: 28S

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Bacillus cereus를 억제하는 Bacillus subtilis HH28의 항균물질 정제와 특성규명 (Purification and Characterization of an Antimicrobial Substance from Bacillus subtilis HH28 Antagonistic to Bacillus cereus)

  • 차현아;정다은;홍성욱;정건섭
    • 한국미생물·생명공학회지
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    • 제42권4호
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    • pp.393-401
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    • 2014
  • 청국장으로부터 Bacillus cereus에 대한 항균활성이 가장 높은 균주를 분리하여 형태학적, 생화학적 특성과 16S rDNA 염기서열 결정을 통해 Bacillus subtilis HH28으로 동정 및 명명하였다. B. subilis HH28의 생육시기에 따른 항균활성을 측정해 본 결과, 생육이 대수증식기인 9시간부터 생성되어 사멸기인 60시간에 가장 높은 활성(80 AU/ml)을 나타내었고 144시간(6일)까지 항균활성을 유지하였다. 항균물질의 정제는 황산암모늄 침전과 DEAE-sepharose fast flow, sephacryl S-200HR를 이용하였고, 19.7배의 정제도와 38.4%의 수율로 정제되었다. Tricine SDS-PAGE와 directed detection을 통해 항균물질의 분자량이 약 3,500 Da임을 알 수 있었다. 또한 본 연구의 항균물질을 B. cereus 뿐만 아니라 Listeria monocytogenes, Vibrio parahaemolyticus의 식중독 균에서도 우수한 항균활성을 나타내었고, 항균작용의 기작은 미생물을 사멸시키는 살균작용이였다. 또한 온도 안정성 실험에서 $40-80^{\circ}C$까지 안정했고, pH 안정성 실험에서는 pH 2-9까지 안정하여 비교적 온도와 pH에 안정하였다. 효소에 대한 영향 실험에서는 단백질 분해효소에 의해 항균활성이 실활되어 본 연구의 항균물질은 단백질성임을 알 수 있었다. 위와 같은 특성으로 보아 B. subtilis HH28이 생산하는 항균물질은 천연 식품 보존제 및 사료 보존제, 항생제 대체 의약품으로 사용할 수 있는 잠재력을 가지고 있으며, 향후 이 항균물질의 정확한 구조 및 특성 규명 등의 연구가 필요하다.

Selection and evaluation of reference genes for gene expression using quantitative real-time PCR in Mythimna separata walker (Lepidoptera: Noctuidae)

  • ZHANG, Bai-Zhong;LIU, Jun-Jie;CHEN, Xi-Ling;YUAN, Guo-Hui
    • Entomological Research
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    • 제48권5호
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    • pp.390-399
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    • 2018
  • In order to precisely assess gene expression levels, the suitable internal reference genes must be served to quantify real-time reverse transcription polymerase chain reaction (RT-qPCR) data. For armyworm, Mythimna separata, which reference genes are suitable for assessing the level of transcriptional expression of target genes have yet to be explored. In this study, eight common reference genes, including ${\beta}$-actin (${\beta}$-ACT), 18 s ribosomal (18S), 28S ribosomal (28S), glyceraldehyde-3-phosphate (GAPDH), elongation fator-alpha ($EF1{\alpha}$), TATA box binding protein (TBP), ribosomal protein L7 (RPL7), and alpha-tubulin (${\alpha}$-TUB) that in different developmental stages, tissues and insecticide treatments of M. separata were evaluated. To further explore whether these genes were suitable to serve as endogenous controls, three software-based approaches (geNorm, BestKeeper, and NormFinder), the delta Ct method, and one web-based comprehensive tool (RefFinder) were employed to analyze and rank the tested genes. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated according to normalized HSP70, and MsepCYP321A10 gene expression data. We found that the most suitable reference genes for the different experimental conditions. For developmental stages, 28S/RPL7 were the optimal reference genes, both $RPL7/EF1{\alpha}$ were suitable for experiments of different tissues, whereas for insecticide treatments, $28S/{\alpha}-TUB$ were suitable for normalizations of expression data. In addition, $28S/{\alpha}-TUB$ were the suitable reference genes because they have the most stable expression among different developmental stages, tissues and insecticide treatments. Our work is the first report on reference gene selection in M. separata, and might serve as a precedent for future gene expression studies.

Streptomyces chromofuscus SMF28을 이용한 Cathepsin B 저해물질의 발효생산 및 특성분석 (Production and Characterization of Cathepsin B Inhibitor from Streptomyces chromofuscus SMF28)

  • 이현숙;김인섭;윤성준;이계준
    • 한국미생물·생명공학회지
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    • 제23권5호
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    • pp.602-608
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    • 1995
  • The aim of the present research program was to construct an optimum fermentation system and to characterize the properties of cathepsin B inhibitor from Streptomyces chromofuscus SMF28. Glucose and casitone were proved to be good carbon source and nitrogen source, respectively. The production of inhibitor was high at lower concentration than 10 mM of inorganic phosphate. The optimum temperature and pH for the production of inhibitor were 30$\circ$C and pH 7, respectively. The production of inhibitor was related to mycelial growth and was affected by medium composition. The inhibitor in culture filtrate of S. chromofuscus SMF28 was purified by butanol extraction, silica gel chromatography, Amberlite IRC-50 (H$^{+}$ form) chromatography, preparative TLC, and preparative HPLC. From amino acid analysis and UV, IR, $^{1}$H-NMR spectroscopic analysis, the inhibitor was identified as a peptide containing valine and phenylalanine derivative.

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능이버섯에서의 lanosterol과 ergosterol의 확인 (Identification of Lanosterol and Ergosterol in Sarcodon aspratus Berk. (S. Ito))

  • 김종봉;박홍덕
    • 생명과학회지
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    • 제10권6호
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    • pp.617-620
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    • 2000
  • Methanol extract of Sarcodon aspratus Berk. (S. Ito) was analyzed by thi layer chromatography, gas chromatography and mass spectrophotometer. Nine fractions from primary methanol extract were observed by TLC. Six fractions were observed by the second TLC analysis of the second and the third fractions. 27,28-digydrolanosterol({TEX}$C_{30}H_{52}${/TEX}O), ergost-7-en-3-ol({TEX}$C_{28}H_{48}${/TEX}O) were identified from two fractions of the second TLC analysis by mass spectrophotometer. The molecular weights of 27,28-dihydrolanosterol and ergost-7-en-3-ol were 413 and 400 respectively.

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제주 연안해수로부터 한천 분해 효소 및 자일란 분해 효소를 생산하는 Catenovulum jejuensis A28-5의 동정 및 특성 규명 (Identification and Characterization of an Agarase- and Xylanse-producing Catenovulum jejuensis A28-5 from Coastal Seawater of Jeju Island, Korea)

  • 김다솜;정가람;배창환;여주홍;지원재
    • 한국미생물·생명공학회지
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    • 제45권2호
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    • pp.168-177
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    • 2017
  • Strain A28-5는대한민국 제주도 연안의 해수샘플로부터 고체배지내 xylan과 agar를 분해하는 균주로 분리되었다. Strain A28-5는그람 음성균으로 한 개의 polar flagella로 운동성을 갖는 $Na^+$ 이온 요구성 균주로 분석되었다. 또한 ampilcillin과 thiostreptone 등의 항생제에 내성을 보였다. Genome 내 G+C content는 43.96%이고, Menaquinone-7 (MK-7)을 predominant quinone으로 함유하고 있었다. Strain A28-5의 세포벽을 구성하는 주요 지방산은 $C_{16:1}$ ${\omega}7c/iso-C_{15:0}$ 2-OH (23.32%), $C_{16:0}$ (21.83%), $C_{18:1}$ ${\omega}7c$ (17.98%)였다. strain A28-5의 16S rRNA gene sequence는 Catenovulum agarivorans YM01와 가장 높은 상동성(98.94%)을 보였으며, Neighbor-Joining phylogenetic tree 제작을 통해서 Catenovulum agarivorans YM01와 가장 높은 근연관계를 보이는 것을 증명하였다. Catenovulum agarivorans YM01과의 DNA-DNA hybridization 분석을 통하여 A28-5을 Catenovulum 속의 신종으로 분류하여Catenovulum jejuensis A28-5로 명명하였다. 이 균주의 액체배양으로부터 준비된 두 종류의 조효소를 이용한 xylan 또는 agarose와의 효소반응액을 Thin layer chromatography로 분석하여 각각 tetramer와 hexamer의 xylooligosaccharides와 (neo)agarooligosacchardes가 생산되는 것을 확인하였다.

Corticotropin-Releasing Hormone (CRH)에 의한 인간 위암 세포(MKN-28)의 Migration 증가 (Enhancement of Cell Migration by Corticotropin-Releasing Hormone (CRH) in Human Gastric Cancer Cell Line, MKN-28)

  • 천소영;조대호
    • IMMUNE NETWORK
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    • 제4권4호
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    • pp.244-249
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    • 2004
  • Background: Corticotropin-Releasing Hormone (CRH), an important regulator of stress response, has a potent immunoregulatory effect with the ability to promote the growth of various cancer through CRH receptor type 1 under stress. Although the metastasized cancers through cell migration are more aggressive than the primary cancers, little is known about the effect of CRH on cell migration. Gastric cancer is prone to metastasize to other tissues and it is reported that gastric cancer is response to various stresses such as oxidative stress. Herein, we studied the relationship between CRH and gastric cancer cell migration. Methods: We used gastric cancer cell line, MKN-28 and tested the CRH receptor type 1 expression on MKN-28 by RT-PCR. To examine the change in the ability of migration by CRH in MKN-28, cells were incubated with CRH and then migration ability was measured using a cell migration assay. Results: We confirmed that CRH receptor type 1 was expressed in MKN-28 and HaCaT cells. The migration ability of MKN-28 cells was increased by CRH in a time-, dose- dependent manner. Conclusion: These data suggest that CRH increases migration ability in gastric cancer cell line and that CRH may be a critical regulator in the metastasis of gastric cancer cell.

삼각형의 변들에 대한 등식을 탐구하는 한 방법에 대한 연구

  • 강인주;한인기
    • East Asian mathematical journal
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    • 제28권2호
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    • pp.197-213
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    • 2012
  • In this paper we study Soltan & Meidman's method that is able to be used in mathematical discovery. We analyze Soltan & Meidman's book "Tozdestva i Neravenstva v Treugolike" that is published in Moldova Republic. In this work we formulate Soltan & Meidman's method related with discovery of triangle's various equalities, and use the method to discovery mathematical equalities. As a result we suggest some new mathematical equalities related with triangle's sides and its proof.

Iron Nutritional Status of Infants and Young Children in the Seoul Area

  • Um, Sung-Sin;Ahn, Hong-Seok;Kim, Soon-Ki;Ha, Jung-Hun
    • Journal of Community Nutrition
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    • 제4권1호
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    • pp.3-11
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    • 2002
  • The purpose of this study is to evaluate the iron nutritional status by investigating dietary intake and analyzing the hematological iron status indices including serum transferrin receptor (sTfR) in 8 to 28 month old infants md young children taking supplementary foods. The nutrient intake of 60 healthy infants and young children from 8 to 24 months of age was investigated by means of a 24-hour recall method, and the subjects were divided into 2 groups (8- 12 months and 13-28 months) according to age. Venous blood samples from these groups were collected and measured for the following : hemoglobin(Hb), hematocrit(Hct) , mean corpuscular volume (MCV), mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration (MCHC), serum ferritin, serum iron, total iron binding capacity (TIBC), and sTfR. Anemia is defined as hemoglobin < 11g /dl , serum ferritin level < 10ng1m1 for iron deficiency , serum transferring receptor(sTfR) > 4.5mg / 1 for iron deficient erythropoiesis. Total daily calorie intake was 934.6 ${\pm}$ 284.5kcal (98.32% of RDA) on average. Average daily iron intake in infants aged 8 to 12 months was 8.92 ${\pm}$ 3.32mg. The mean daily iron intake in infants aged 13 to 28 months was 7.15 ${\pm}$ 3.35mg (90% of Recommended Dietary Allowance, RDA). Mean values for Hb, Hct sew ferritin and sTfR were 12.10 ${\pm}$ 0.77g141,36.02 ${\pm}$ 2.31%,20.91 ${\pm}$ 11.58ng/m1 and 3.78 ${\pm}$ 1.47mg /1, respectively. In the young children from 13 to 28 months of age, the prevalence of anemia was 5.6%. The prevalence of iron deficiency was 9.5% in those from 8 to 12 months of age, and 27.8% in those from 13 to 28 months of age. The prevalence of iron deficient erythropoiesis was 16.7% in infants aged 8 to 12 months and 44.4% in those aged 13 to 28 months. The prevalence of both serum ferritin level < 10ng/m1 sTfR > 4.5mg/1 was 22% in the young children aged 13 to 28 months. The measureand ment of sTfR may be a promising new tool in diagnosis of iron deficiency in early childhood when the iron deficiency is prevalent. It seems appropriate to emphasize nutritional education and evaluation to promote the iron nutritional status of infants and young children.