• 제목/요약/키워드: Gene Identification

검색결과 1,724건 처리시간 0.024초

염소의 베타-락토글로불린 유전자 프로모터의 음성 조절 인자 규명 (Identification of the Negative Regulatory Element on the Caprine $\beta$ Lactoglobulin Promoter)

  • 김재만;유명희
    • 한국동물학회지
    • /
    • 제38권3호
    • /
    • pp.433-441
    • /
    • 1995
  • 염소 베타-락토글로불린 유전자 프로모터의 유선 조직 특이성은, 비 발현 세포에서 -470 에서 -205 부위에 의해서 매개되는 억제 조절에 의해서 보증된다. 이 음성 조절 기작과 억제 조절을 매개하는 인자를 확인하기 위하여 상류 염기 서열을 자세히 분석하였다. 상류 염기 서열은 어느 방향으로 위치하든 연결된 베타-락토글로불린 유전자 프로모터의 활성을 억제할 수 있었다. 이와 같이 규명된 염소 베타-락토글로불린 유전자의 잠정적 음성 조절 인자는 다른 유전자의 프로모터들에 대해서는 다양한 활성을 보였는데, herpes simplex 바이러스의 thimidine kinase 프로모터는 상류 염기서열의 방향에 따라 억제 또는 활성화되었으며, SV40 프로모터는 억제되기보다는 오히려 활성화되었다. 염소 베타-락토글로불린 유전자의 억제 조절 인자를 포함하는 조절 부위는 비 유선 세포인 HeLa 및 CV-1 세포에서 추출된 핵 추출물에 의해서 이동성이 강력하게 지연되는 반면, 유선세포인 HC11 세포의 핵 추출물에 의해서는 약하게 지연되었다. 이와 같은 활성 억제와 인자 결합간의 연관성은 foot-printing 분석에서 관찰된 결합 부위가 염소의 베타-락토글로불린 유전자의 조직특이적 억제에 작용할 음성 조절 인자일 가능성을 시사한다.

  • PDF

Cytokine Reporter Mouse System for Screening Novel IL12/23 p40-inducing Compounds

  • Im, Wooseok;Kim, Hyojeong;Yun, Daesun;Seo, Sung-Yum;Park, Se-Ho;Locksley, Richard M.;Hong, Seokmann
    • Molecules and Cells
    • /
    • 제20권2호
    • /
    • pp.288-296
    • /
    • 2005
  • Cytokines interleukin (IL) 12 and 23 play critical roles in linking innate and adaptive immune responses. They are members of heterodimeric cytokines, sharing a subunit p40. Although IL12/23 p40 is mainly induced in macrophages and dendritic cells (DCs) after stimulation with microbial Toll-like receptor ligands, methods to monitor the cells that produce IL12/23 p40 in vivo are limited. Recently, the mouse model to track p40-expressing cells with fluorescent reporter, yellow fluorescent protein, has been developed. Macrophages and DCs from these mice faithfully reported p40 induction using the fluorescent marker. Here we took advantage of these reporter mice to screen bio-compounds for p40-inducing activity. After screening hundreds of compounds, we found several extracts inducing IL12/23 p40 gene expression. Treatment of DCs with these extracts induced the expression of MHC class II and co-stimulatory molecules, which implies that these might be useful as adjuvants. Next, the in vivo target immune cells of candidate compounds were examined. The reporter system can be useful to identify cells producing IL12 or IL23 in vivo as well as in vitro. Thus, our cytokine reporter system proved to be a valuable reagent for screening for immunostimulatory molecules and identification of target cells in vivo.

Isolation and Identification of Lactobacillus kimchicus sp.nov and Bioconversion of Ginseng Saponin

  • Liang, Zhi-Qi;Kim, Ho-Bin;Kim, Yeon-Ju;Wang, Hong-Tao;Yang, Deok-Chun
    • 한국자원식물학회:학술대회논문집
    • /
    • 한국자원식물학회 2010년도 정기총회 및 추계학술발표회
    • /
    • pp.15-15
    • /
    • 2010
  • Ginseng contained many different kinds of saponin which was the most valuable for people, but its yield cannot satisfy the demand using traditional extract methods. Enzyme transformation is a conformable and highly performed method which was fit for today. A ${\beta}$-glucosidase producing bacterium ($DCY51^T$) was isolated from Korean fermented-vegetable food kimchi. The 16S rRNA gene sequence analysis revealed that the strain $DCY51^T$ belongs to the genus Lactobacillus. The highest sequence similarity was found with Lactobacillus paracollinoides LMG $22473^T$ and Lactobacillus collinoides LMG $9194^T$ with levels of 16S rDNA similarity of 97.4% and 97.3%, respectively. Based on the above results the strain $DCY51^T$ placed in the genus Lactobacillus and proposed a new species, Lactobacillus kimchicus sp. nov. $DCY51^T$ (= KCTC $12976^T$ = JCM $15530^T$). It was culture solution reacted with Red Ginseng extract and $Rb_1$, respectively. The medium of bacteria was the liquid of MRS, the temperatures of growing and reacting between bacteria liquid and saponin were samely $37^{\circ}C$, there spective reacting time were 12 hours and 48 hours. Thus we got different saponins, and TLC and HPLC analysis showed that: enzyme respectively reacted with $Rb_1$ and Red Ginseng extract got the transformed saponin, respectively. The polarity position in TLC was a little higher than Rd; and the polarity position was the same as that of Compound K's, the saponin obtained from HPLC and other experimental results was not Compound K. The constitution of its saponin was hoped to be further confirmed.

  • PDF

Identification of Coccoidal Bacteria in Traditional Fermented Milk Products from Mongolia, and the Fermentation Properties of the Predominant Species, Streptococcus thermophilus

  • Ren, Yan;Liu, Wenjun;Zhang, Heping
    • 한국축산식품학회지
    • /
    • 제35권5호
    • /
    • pp.683-691
    • /
    • 2015
  • The objective of this study was to identify the coccoidal bacteria present in 188 samples of fermented yaks’, mares’ and cows’ milk products collected from 12 different regions in Mongolia. Furthermore, we evaluated the fermentation properties of ten selected isolates of the predominant species, Streptococcus (S.) thermophiles, during the process of milk fermentation and subsequent storage of the resulting yoghurt at 4℃. Overall, 159 isolates were obtained from 188 samples using M17 agar. These isolates were presumed to be lactic acid bacteria based on their gram-positive and catalase-negative properties, and were identified to species level using 16S rRNA gene sequence analysis. These coccoid isolates were distributed in four genera and six species: Enterococcus (E.) durans, Enterococcus (E.) faecalis, Lactococcus (Lac.) subsp. lactis, Leuconostoc (Leuc.) lactis, Leuconostoc (Leuc.) mesenteroides. subsp. mesenteroides and S. thermophilus. Among these S. thermophilus was the most common species in most samples. From evaluation of the fermentation characteristics (viable counts, pH, titratable acidity [TA]) of ten selected S. thermophilus isolates we could identify four isolates (IMAU 20246, IMAU20764, IMAU20729 and IMAU20738) that were fast acid producers. IMAU20246 produced the highest concentrations of lactic acid and formic acid. These isolates have potential as starter cultures for yoghurt production.

Genetic and Phenotypic Diversity of Plant Growth Promoting Rhizobacteria Isolated from Sugarcane Plants Growing in Pakistan

  • Mehnaz, Samina;Baig, Deeba N.;Lazarovits, George
    • Journal of Microbiology and Biotechnology
    • /
    • 제20권12호
    • /
    • pp.1614-1623
    • /
    • 2010
  • Bacteria were isolated from roots of sugarcane varieties grown in the fields of Punjab. They were identified by using API20E/NE bacterial identification kits and from sequences of 16S rRNA and amplicons of the cpn60 gene. The majority of bacteria were found to belong to the genera of Enterobacter, Pseudomonas, and Klebsiella, but members of genera Azospirillum, Rhizobium, Rahnella, Delftia, Caulobacter, Pannonibacter, Xanthomonas, and Stenotrophomonas were also found. The community, however, was dominated by members of the Pseudomonadaceae and Enterobacteriaceae, as representatives of these genera were found in samples from every variety and location examined. All isolates were tested for the presence of five enzymes and seven factors known to be associated with plant growth promotion. Ten isolates showed lipase activity and eight were positive for protease activity. Cellulase, chitinase, and pectinase were not detected in any strain. Nine strains showed nitrogen fixing ability (acetylene reduction assay) and 26 were capable of solubilizing phosphate. In the presence of 100 mg/l tryptophan, all strains except one produced indole acetic acid in the growth medium. All isolates were positive for ACC deaminase activity. Six strains produced homoserine lactones and three produced HCN and hexamate type siderophores. One isolate was capable of inhibiting the growth of 24 pathogenic fungal strains of Colletotrichum, Fusarium, Pythium, and Rhizoctonia spp. In tests of their abilities to grow under a range of temperature, pH, and NaCl concentrations, all isolates grew well on plates with 3% NaCl and most of them grew well at 4 to $41^{\circ}C$ and at pH 11.

The ACC deaminase from rhizobateria promoted resistance of salininty stress in seedling and growth of plant

  • Soh, Byoung-Yul;Lee, Gun-Woong;Ju, Jae-Eun;Kim, Hae-Min;Chae, Jong-Chan;Lee, Yong-Hoon;Oh, Byung-Taek;Lee, Kui-Jae
    • 한국자원식물학회:학술대회논문집
    • /
    • 한국자원식물학회 2010년도 정기총회 및 추계학술발표회
    • /
    • pp.17-17
    • /
    • 2010
  • Rhizobacteria are a diverse group of free-living soil bacteria that live in plant rhizosphere and colonized the root system. Plant growth-promoting rhizobacteria (PGPR) possessing ACC deaminase (ACCD) can reduce ACC and ethylene in plant tissue and mediated the growth of plants under various stresses including salt stress. ACCD decrease ethylene levels in plant tissue that produce high levels of ethylene in tissue via elevated levels of ACC under salt stress. We selected strains of Pseudomonas sp. possessing ACCD activity for their ability to promote plant growth under salt stress from soil sample collected at Byeonsan, Jeonbuk, South Korea. The Pseudomonas strains possessing ACCD increased the rate of the seedling and growth of chinese cabbage seeds under salt stress. We cloned ACCD gene from P.fluorescens and expressed recombinant protein in Escherichia coli. The active form of recombinant ACCD converted ACC to a-ketobutyrate. The in vivo treatment of recombinant ACCD itself increase the rate of the seedling and growth of Chinese cabbage seeds under salt stress. The polyclonal P.fluorescens anti-ACCD antibody specifically reacted with ACCD originated from Pseudomonas. This indicates that the antibody might act as an important indicator for ACCD driven from Pseudomonas exhibiting plant growth-promoting activity. This study will be useful for identification of newly isolated PGPR containing ACCD and exploioting the ACCD activity from PGPR against various biotic and abiotic stresses.

  • PDF

Genetic Variations of Trichophyton rubrum Clinical Isolates from Korea

  • Yoon, Nam-Sup;Kim, Hyunjung;Park, Sung-Bae;Park, Min;Kim, Sunghyun;Kim, Young-Kwon
    • 대한의생명과학회지
    • /
    • 제24권3호
    • /
    • pp.221-229
    • /
    • 2018
  • Trichophyton rubrum is one of the well-known pathogenic fungi and causes dermatophytosis and cutaneous mycosis in human world widely. However, there are not an available sequence type (ST) classification methods and previous studies for T. rubrum until now. Therefore, currently, molecular biological tools using their DNA sequences are used for genotype identification and classification. In the present study, in order to characterize the genetic diversity and the phylogenetic relation of T. rubrum clinical isolates, five different housekeeping genes, such as actin (ACT), calmodulin (CAL), RNA polymerase II (RPB2), superoxide dismutase 2 (SOD2), and ${\beta}$-tubulin (BT2) were analyzed using by multilocus sequence typing (MLST). Also, DNA sequence analysis was performed to examine the differences between the sequences of Trichophyton strains and the identified genetic variations sequence. As a result, most of the sequences were shown to have highly matched rates in their housekeeping genes. However, genetic variations were found on three different positions of ${\beta}$-tubulin gene and were shown to have changed from $C{\rightarrow}G$ (1766), $G{\rightarrow}T$ (1876), and $C{\rightarrow}A$ (1886). To confirm the association with T. rubrum inheritance, a phylogenetic tree analysis was performed. It was classified as four clusters, but there was little significant correlation. Even so, MLST analysis is believed to be helpful for determining the genetic variations of T. rubrum in cases where there is more large-scale data accumulation. In conclusion, the present study demonstrated the first MLST analysis of T. rubrum in Korea and explored the possibility that MLST could be a useful tool for studying the epidemiology and evolution of T. rubrum through further studies.

Identification and Antibiotic Susceptibility of the Bacteria from Non-odontogenic Infectious Lesions

  • Kim, Yong Min;Kim, Jae-Jin;Kim, Mija;Park, Soon-Nang;Kim, Hwa-Sook;Kook, Joong-Ki;Kim, Hak Kyun
    • International Journal of Oral Biology
    • /
    • 제39권2호
    • /
    • pp.87-95
    • /
    • 2014
  • The purpose of this study was to isolate and identify bacteria from the 4 patients with non-odontogenic infectious lesions (mucormycosis, chronic inflammation from wound infection, and two actinomycosis) and determine their antimicrobial susceptibility against eight antibiotics. Bacterial culture was performed under three culture conditions (anaerobic, $CO_2$, and aerobic incubator). The bacterial strains were identified by 16S rRNA gene (16S rDNA) sequence comparison analysis method. For investigating the antimicrobial susceptibility of the bacteria against eight antibiotics, penicillin G, amoxicillin, tetracycline, cefuroxime, erythromycin, clindamycin, vancomycin, and Augmentin$^{(R)}$ (amoxicillin + clavulanic acid), minimum inhibitory concentration (MIC) measurement was performed using broth microdilution assay. Nosocomial pathogens such as Enterococcus faecalis, Klebsiella pneumoniae, Bacillus subtilis, and Neisseria flavescens were isolated from mucormycosis. Veillonella parvula, Enterobacter hormaechei, and Acinetobacter calcoaceticus were isolated from chronic inflammatory lesion. Actinomyces massiliensis was isolated from actinomycosis in parotid gland. Capnocytophaga ochracea was isolated from actinomycosis in buccal region in anaerobic condition. There was no susceptible antibiotic to all bacteria in mucormycosis. Tetracycline was susceptible to all bacteria in chronic inflammation. C. ochracea was resistant to vancomycin and penicillin G; and other antibiotics showed susceptibility to all bacteria in actinomycosis. The results indicated that the combined treatment of two or more antibiotics is better than single antibiotic treatment in mucormycosis, and penicillin is the first recommended antibiotic to treat actinomycosis.

옥수수 이삭썩음병에 관여하는 Fusarium속균의 동정 (Identification of Fusarium Species Associated with Corn Ear Rot)

  • 최효원;김정미;김진희;홍성기;김완규;천세철
    • 한국균학회지
    • /
    • 제37권2호
    • /
    • pp.121-129
    • /
    • 2009
  • In 2007, a total of 77 isolates of Fusarium spp. were obtained from ear rot symptoms of corns collected from 5 locations in Gangwon Province, Korea. The fungal isolates were identified based on their morphological features. Out of the isolates, fifteen isolates were identified as Fusarium verticillioides which formed microconidia in long chains on monophialides. Four isolates were identified as F. subglutinans which formed microconida only on false heads. Six isolates were identified as F. graminearum which produced red pigment in PDA culture. Besides these Fusarium species, F. napiform, F. nygamai, and F. oxysporum were identified from the rest isolates. To assess for genetic diversity of the isolates, a random amplified polymorphic DNA(RAPD) technique was carried out using URP primers. The results from the RAPD analysis showed that the isolates from corn were divided into 6 groups. These RAPD groups of the Fusarium species corresponded to morphological characters of the Fusarium species. The phylogenetic analysis of most isolates by DNA sequencing of EF-1$\alpha$ gene corresponded to morphological characters of the Fusarium species. The results of pathogenicity tests by two inoculation methods revealed that F. verticillioides, F. graminearum and F. subglutinans are strongly pathogenic to corn stalks.

Burkholderia pyrrocinia LA101 선발 (Selection of Burkholderia pyrrocinia LA101)

  • 나정우;장명준;안승원;박윤진;조용구
    • 한국환경과학회지
    • /
    • 제29권5호
    • /
    • pp.435-443
    • /
    • 2020
  • The purpose of this study was to investigate eco-friendly measures to manage major diseases which cause heavy economic damages to ginseng. Morphological, physicochemical, and molecular biological species identification was carried out after isolating useful antagonistic bacteria from ginseng fields. In addition, optimal conditions for mass culture were established, and he efficacy of the bacteria in the prevention of the diseases was verified in the field. The results showed that about 150 bacteria were extracted from 150 ginseng fields in the whole county. Among them, B. pyrrocinia LA101 was finally selected, which had a strong antagonistic potency against Alternaria panax, Botrytis cinerea, Rhizoctonia solani, and Cylindrocarpon destructans on agar media. The B. pyrrocinia LA101 is a baculiform gram-negative bacterium identified as Burkholderia pyrrocinia according to results from an API(Analytical Profile Index) kit, 16S rRNA, and gyrase gene sequencing analysis. It was donated to the microbe bank of the Agricultural Genetic Resources Center at the National Academy of Agriculture Science under the Rural Development Administration on September 28, 2011 (Donation No. KACC91663P). A patent for the mass culture technology was granted in August 2012 (Patent No. 10-1175532).