• Title/Summary/Keyword: 5S rRNA genes

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Reevaluation of Enumeration of Bacillus cereus Grown on Mannitol-Egg York-Polymyxin B Agar (Mannitol-Egg York-Polymyxin B 선택 배지에서 Bacillus cereus 계수 방법의 재평가)

  • Yun, Suk-Hyun;Kim, Yong-Sang;Jeong, Do-Yeon;Hahn, Kum-Su;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.208-214
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    • 2009
  • To avoid ambiguity in counting the number of colony, about 1,500 of colonies grown on B. cereus selective agar plates were grouped into 12 types by morphological difference and then identified by biochemical and 16S rDNA nucleotide sequence. Among them, seven colony types with 11 to 15 mm diameters of halo were identified as B. cereus or B. cereus subsp. cytotoxis. Five mm sized colonies with no halo, which have not been considered as B. cereus according to the manufacturer's manual, were identified as B. cereus. A colony type with double halos of only 6 mm in diameter was also B. cereus. Other three types were proven to be Enterococcus sp., Brevibacillus sp., and B. subtilis, respectively. PCR results showed that only 9 types that are identified as B. cereus strains harbor at least one of B. cereus toxin genes.

Seasonal Variations in the Bacterial Community of Gwangyang Bay Seawater (광양만 해수의 세균 군집의 계절적 변화)

  • Park, Seong Chan;Lee, Ji Hee;Kang, Joo Won;Baik, Keun Sik;Seong, Chi Nam
    • Journal of Life Science
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    • v.24 no.5
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    • pp.522-531
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    • 2014
  • Seasonal variations in the bacterial community of Gwangyang Bay seawater were analyzed using both isolation and cultivation-independent methods. Amplified rDNA restriction analysis was applied to 200 bacterial isolates. Bacterial isolates were composed of four phyla: Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes. Pyrosequencing was conducted, in addition to denaturing gradient gel electrophoresis (DGGE) of genomic DNA extracted directly from the water samples. The bacterial sequences obtained by pyrosequencing of 16S rRNA genes consisted of 24 phyla in the spring and summer, 39 in the fall, and 32 in the winter. The diversity index was high in the fall, whereas the dominancy index was high in the spring. In the spring, phylum Firmicutes was dominant, whereas phylum Proteobacteria dominated in the other three seasons. The second most dominant phyla were Proteobacteria in the spring, Firmicutes in the summer, and Bacteroidetes both in the fall and winter. Bacilliaceae was the most predominant family in the spring. Rhodobacteraceae and Bacilliaceae dominated in the summer, and Rhodobacteraceae dominated in the winter. Neither was dominant in the fall Twenty-seven bands purified from DGGE profiles were cloned and analyzed phylogenetically. In the spring, phylum Firmicutes dominated, followed by Proteobacteria. Proteobacteria dominated in all other seasons. Thus, two cultivation-independent methods for determination of seasonal variation patterns at the phylum level were in accordance with each other.

Identification of Differentially Expressed Genes in Four Different Growing Stages in Korea Native Chicken Liver (황갈색 재래닭의 간에서 성장 단계별 차등 발현 유전자 분석)

  • Lee, K.Y.;Yu, S.L.;Jung, K.C.;Jang, B.K.;Choi, K.D.;Lee, J.H.
    • Korean Journal of Poultry Science
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    • v.34 no.2
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    • pp.85-90
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    • 2007
  • The chicken liver has been involved in various biological functions including detoxification, glycogen storage and plasma protein synthesis. The aim of this study was to investigate differentially expressed genes in chicken liver in four different growing stages. Using 10 arbitrary Annealing Control Primers (ACPs), five differentially expressed genes have been identified. Based on the Basic Local Alignment Search Tool (BLAST) search results, three of them were matched with previously known genes, and the other two were matched with unknown EST sequence and a hypothetical protein, respectively. In order to confirm the expression results, quantitative real-time PCR was also performed. The high similarities between the expression data using arbitrary ACPs and quantitative real-time PCR indicate that the identified genes are the real differentially expressed genes in different growing stages. The genes identified in this study can be used as valuable biomarkers in chicken with further investigation of the functions.

Mycolic Acid-Containing Actinomycetes Associated with Activated Sludge Foam

  • Seong, Chi-Nam;Kim, Young-Sook;Baik, Ken-Shik;Lee, Soon-Dong;Hah, Yung-Chil;Kim, Seung-Bum;Goodfellow, Michael
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.66-72
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    • 1999
  • Mycolic acid-containing actinomycetes associated with extensive foaming in the aeration basin of the activated sludge process were isolated and analyzed by phenotypical, chemotaxonomical and phylogenetic methods. Whole cell sugar patterns of two isolates were pattern A. The nearly complete sequences of the 16S rRNA genes (rDNAs) of the isolates were determined and compared by using several tree-making algorithms. With polyphasic methods, strain SCNU1 was identified as Gordona sputi, and strain SCNU5 assigned to the genus Tsukamurella. The presence of opportunistic pathogens of chronic lung infections within foams can cause public health problems and render waste-treatment processes inefficient.

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Raoultella ornithinolytica as a Potential Candidate for Bioremediation of Heavy Metal from Contaminated Environments

  • Laila Ibrahim Faqe Salih;Rezan Omer Rasheed;Sirwan Muhsin Muhammed
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.895-908
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    • 2023
  • Disposal of waste containing heavy metals into the environment is a major threat to human health and can result in toxic or chronic poisoning in aquatic life. In the current study, metal-resistant Raoultella ornithinolytica was isolated from metal-contaminated samples collected from the Tanjaro River, located southwest of Sulaymaniyah, Iraq. R. ornithinolytica was identified by partial amplification of 16S rRNA. The uptake potency of heavy metals was assessed using inductively coupled plasma-optical emission spectroscopy (ICP-OES) and indicated that R. ornithinolytica removed 67, 89, 63.4, 55.6, 56.5, 65, and 61.9% of Cd, Pb, Cr, Ni, Zn, Co, and Fe, respectively. These removal rates were influenced by temperature, pH, and contact time; at 35℃ and pH 5 with a change in the incubation time, the reduction rate improved from 89 to 95% for Pb, from 36.4 to 45% for Cu, and from 55.6 to 64% for Ni. Gene analysis indicated that R. ornithinolytica contained pbrT, chrB, nccA, iroN, and czcA genes, but the pcoD gene was absent. Energy-dispersive X-ray spectroscopy (EDS) images showed evidence of metal ion binding on the cell wall surface with different rates of binding. Transmission electron microscopy (TEM) detected different mechanisms for metal particle localization; cell surface adsorption was the main mechanism for Pb, Zn, and Co uptake, while Cd, Ni, and Fe were accumulated inside the cell. The current study describes, for the first time, the isolation of R. ornithinolytica from metal-contaminated water, which can be used as an eco-friendly biological expedient for the remediation and detoxification of metals from contaminated environments.

Effects of Dohongsamul-Tang on the Gene Expression of Photothrombotic Ischemia Mouse Model (도홍사물탕(桃紅四物湯)이 광화학적 뇌경색 마우스의 유전자 발현에 미치는 영향)

  • Cho, Kwon-Il;Kim, Hye-Yoon;Ko, Seok-Jae;Lee, Seong-Geun;Shin, Sun-Ho;Moon, Byung-Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.3
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    • pp.645-661
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    • 2009
  • The water extract of Dohongsamul-Tang(DHSMT) has been traditionally used to stroke and brain injuries in Oriental Medicine. The present study was designed to investigate the effects of DHSMT on the gene expression profile of cerebral infarction by cDNA microarray in photothrombotic ischemia mouse model. Photothrombotic ischemia was induced in stereotactically held male BALB/c mice using rose bengal and cold light. MRI was performed 24 hours after inducing photothrombosis using 1.5 T MRI and 47 mm surface coil to obtain T2-weighted, and contrast-enhanced images. After MRI test, animal was sacrificed and the brain sections were stained for hematoxylin and eosin and immunohistochemistry. MRI and histological analysis revealed that lesion of thrombotic ischemia was well induced in the cortex with the evidence of biological courses of infarction. The target area of thrombotic infarction was 1 mm anterior to bregma and 3 mm lateral to midline with 2 mm in diameter, which were decreased by administration of DHSMT. To assess gene expression pattern of cerebral infarction, mRNA was isolated and reacted with microarray chip(Agilant's DNA Microarray 44K). Scatter and MA plot analysis were performed to clustering of each functional genes. M value [M=log2(R/G), A={log2(R ${\times}$ G)}/2] was between -0.5 and +0.5 with 40% difference. After pretreatment with DHSMT, the expression levels of mRNA of many genes involved in various signaling pathway such as apoptosis, cell cycle, cell proliferation, response to oxidative stress, immune response, angiogenesis, and inflammatory cytokine were markedly inhibited in photothrombotic ischemia lesion compared to the control group. These results suggest that DHSMT prevent ischemic death of brain on photothrombotic ischemia model of mice through modulation of gene expression at the transcriptional level.

Isolation of Mannanase-producing Bacteria, Bacillus subtilis WL-6 and WL-11, and Cloning and Characterization of Mannanase (Bacillus subtilis 분리균 2주 유래 mannanases의 특성 비교)

  • Yoon, Ki-Hong
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1113-1120
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    • 2016
  • Two bacterial strains producing extracellular man nanase were isolated from doenjang, a traditionally fermented soybean paste in Korea. The isolates, WL-6 and WL-11, were identified as Bacillus subtiis on the basis of their 16S rRNA gene sequences, morphological, and biochemical properties. Two genes encoding the mannanase of both B. subtilis WL-6 and B. subtilis WL-11 were each cloned into Escherichia coli, and their nucleotide sequences were determined. Both mannanase genes consisted of 1,086 nucleotides, encoding polypeptides of 362 amino acid residues. The deduced amino acid sequences of the two WL-6 and WL-11 mannanases, designated Man6 and Man11, respectively, differed from each other by eight amino acid residues, and they were highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. The 26 amino acid stretch in the N-terminus of Man6 and Man11 was a predicted signal peptide. Both Man6 and Man11 were localized at the level of 94–95% in an intracellular fraction of recombinant E. coli cells. The enzymes hydrolyzed both locust bean gum and mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, forming mannobiose and mannotriose as predominant products. The optimal reaction conditions were 55°C and pH 6.0 for Man6, and 60°C and pH 5.5 for Man11. Man11 was more stable than Man6 at high temperatures.

Cold-Seep Sediment Harbors Phylogenetically Diverse Uncultured Bacteria

  • Cho, Jae-Chang;Lee, Sang-Hoon;Oh, Hae-Ryun;Lee, Jung-Hyun;Kim, Sang-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.906-913
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    • 2004
  • A culture-independent molecular phylogenetic survey was carried out on the bacterial community in cold-seep sediment at Edison Seamount, south of Lihir Island, Papua New Guinea. Small-subunit rRNA genes were amplified directly from the sediment DNA by PCR and cloned. The majority of the cloned 16S rRNA gene sequences were most closely related to as-yet-uncultivated microorganisms found in deep-sea sediments, and were primarily affiliated with one of four groups: the $\gamma$-, $\delta$-, and $\epsilon$-subdivisions of Proteobacteria, and Cytophaga-Flavobacterium-Bacteroides. We did not recover any sequences related to cyanobacteria, prochlorophytes, and $\alpha$-Proteobacteria, which are known to occur in great abundance within the surface mixed layer of the Atlantic and Pacific Oceans. The majority of the cloned $\gamma$-and $\epsilon$-Proteobacterial sequences were closely related to chemoautotrophic sulfur-oxidizing symbionts of marine benthic fauna, and the $\delta$-Proteobacterial sequences to sulfate- and sulfur-reducing bacteria, indicating that they might play an important role in chemoautotrophic primary production and the sulfur cycle in the cold-seep area. There results demonstrate the high diversity of the bacterial community in the cold-seep sediment, and substantially expand knowledge of the extent of bacterial diversity in this formidable and unique habitat.

Effect of Korean pine nut oil on hepatic iron, copper, and zinc status and expression of genes and proteins related to iron absorption in diet-induced obese mice

  • Shin, Sunhye;Lim, Yeseo;Chung, Jayong;Park, Soyoung;Han, Sung Nim
    • Journal of Nutrition and Health
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    • v.54 no.5
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    • pp.435-447
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    • 2021
  • Purpose: Body adiposity is negatively correlated with hepatic iron status, and Korean pine nut oil (PNO) has been reported to reduce adiposity. Therefore, we aimed to study the effects of PNO on adiposity, hepatic mineral status, and the expression of genes and proteins involved in iron absorption. Methods: Five-week-old male C57BL/6 mice were fed a control diet containing 10% kcal from PNO (PC) or soybean oil (SBO; SC), or a high-fat diet (HFD) containing 35% kcal from lard and 10% kcal from PNO (PHFD) or SBO (SHFD). Hepatic iron, copper, and zinc content; and expression of genes and proteins related to iron absorption were measured. Results: HFD-fed mice had a higher white fat mass (2-fold; p < 0.001), lower hepatic iron content (25% lower; p < 0.001), and lower hepatic Hamp (p = 0.028) and duodenal Dcytb mRNA levels (p = 0.037) compared to the control diet-fed mice. Hepatic iron status was negatively correlated with body weight (r = -0.607, p < 0.001) and white fat mass (r = -0.745, p < 0.001). Although the PHFD group gained less body weight (18% less; p < 0.05) and white fat mass (18% less; p < 0.05) than the SHFD group, the hepatic iron status impaired by the HFD feeding did not improve. The expression of hepatic and duodenal ferroportin protein was not affected by the fat amount or the oil type. PNO-fed mice had significantly lower Slc11a2 (p = 0.022) and Slc40a1 expression (p = 0.027) compared to SBO-fed mice. However, the PC group had a higher Heph expression than the SC group (p < 0.05). The hepatic copper and zinc content did not differ between the four diet groups, but hepatic copper content adjusted by body weight was significantly lower in the HFD-fed mice compared to the control diet-fed mice. Conclusion: HFD-induced obesity decreased hepatic iron storage by affecting the regulation of genes related to iron absorption; however, the 18% less white fat mass in the PHFD group was not enough to improve the iron status compared to the SHFD group. The hepatic copper and zinc status was not altered by the fat amount or the oil type.

Microbial Diversity in the Enrichment Cultures from the Fermented Beverage of Plant Extract Using Ribosomal RNA Sequence Analysis (라이보좀 RNA 염기서열 분석을 이용한 집식배양된 식물추출물발효음료의 미생물 다양성)

  • Lee, Choung Kyu;Kim, Baolo;Kang, Young Min;Lee, Hee Yul;Hwang, Chung Eun;Ahn, Min Ju;Seo, Weon Taek;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.351-359
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    • 2014
  • A beverage was produced by the fermentation of mixed extracts from the various fruits, vegetables, algae, and medical herbs. The physicochemical properties of the fermented beverage of plant extracts (FBPE) and microbial diversity were analyzed in cultures enriched from FBPE using 16S and 26S rRNA gene sequence analyses. The pH, acidity, $^{\circ}brix$, reducing sugar, and alcohol contents of the FBPE were determined to be the 3.48, 1.68%, 70.0, 1,026 g/L, and 3.5%, respectively. The most abundant free sugar and organic acid in the FBPE were glucose (567.83 g/L) and tartaric acid (93.68 mg/L), respectively. Lactobacillus homohiochii was the predominant species in all enriched culture samples: 100% of the species in 0B (0% sugar) and 40B (40% sugar) libraries and 95.6% of 20B library (20% sugar). Lactobacillus fructivorans was detected in the 20B library. The predominant species in the samples of enrichment cultures collected from FBPE with three different sugar concentrations were: Candida zeylanoides (45.2%) in the 0Y library (0% sugar), Candida lactis-condensi (35.7%) and C. zeylanoides (35.7%) in the 20Y library (20% sugar), and C. lactis-condensi (38.1%) in the 40Y library (40% sugar). This result may provide a useful frame of reference for further analyses of microbial population dynamics in FBPE.