• Title/Summary/Keyword: ITS regions of rDNA

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Demethylation of CpG islands in the 5' upstream regions mediates the expression of the human testis-specific gene MAGEB16 and its mouse homolog Mageb16

  • Liu, Yunqiang;Wang, Meiling;Jiang, Siyuan;Lu, Yongjie;Tao, Dachang;Yang, Yuan;Ma, Yongxin;Zhang, Sizhong
    • BMB Reports
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    • 제47권2호
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    • pp.86-91
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    • 2014
  • Tissue-specific gene expression is regulated by epigenetic modification involving trans-acting factors. Here, we identified that the human MAGEB16 gene and its mouse homolog, Mageb16, are only expressed in the testis. To investigate the mechanism governing their expression, the promoter methylation status of these genes was examined in different samples. Two CpG islands (CGIs) in the 5' upstream region of MAGEB16 were highly demethylated in human testes, whereas they were methylated in cells without MAGEB16 expression. Similarly, the CGI in Mageb16 was hypomethylated in mouse testes but hypermethylated in other tissues and cells without Mageb16 expression. Additionally, the expression of these genes could be activated by treatment with the demethylation agent 5'-aza-2'-deoxycytidine (5'-aza-CdR). Luciferase assays revealed that both gene promoter activities were inhibited by methylation of the CGI regions. Therefore, we propose that the testis-specific expression of MAGEB16 and Mageb16 is regulated by the methylation status of their promoter regions.

Growth Promotion of Pepper Plants by Pantoea ananatis B1-9 and its Efficient Endophytic Colonization Capacity in Plant Tissues

  • Kim, Su-Nam;Cho, Won-Kyong;Kim, Won-Il;Jee, Hyeong-Jin;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • 제28권3호
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    • pp.270-281
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    • 2012
  • The bacteria B1-9 that was isolated from the rhizosphere of the green onion could promote growth of pepper, cucumber, tomato, and melon plants. In particular, pepper yield after B1-9 treatment on the seedling was increased about 3 times higher than that of control plants in a field experiment. Partial 16S rDNA sequences revealed that B1-9 belongs to the genus Pantoea ananatis. Pathogenecity tests showed non-pathogenic on kimchi cabbage, carrot, and onion. The functional characterization study demonstrated B1-9's ability to function in phosphate solubilization, sulfur oxidation, nitrogen fixation, and indole-3-acetic acid production. To trace colonization patterns of B1-9 in pepper plant tissues, we used $DRAQ5^{TM}$ fluorescent dye, which stains the DNAs of bacteria and plant cells. A large number of B1-9 cells were found on the surfaces of roots and stems as well as in guard cells. Furthermore, several colonized B1-9 cells resided in inner cortical plant cells. Treatment of rhizosphere regions with strain B1-9 can result in efficient colonization of plants and promote plant growth from the seedling to mature plant stage. In summary, strain B1-9 can be successfully applied in the pepper plantation because of its high colonization capacity in plant tissues, as well as properties that promote efficient plant growth.

Molecular Cloning and Analysis of the Gene for P-450 Hydroxylase from Pseudonocardia autotrophica IFO 12743

  • Kim, Jung-Mee;Younmie Jin;Hyun, Chang-Gu;Kim, Jong-Hee;Lee, Hong-Sub;Kang, Dae-Kyung;Kang, Dae-Jung;Kim, Tae-Yong;Suh, Joo-Won
    • Journal of Microbiology
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    • 제40권3호
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    • pp.211-218
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    • 2002
  • A 4.8-kb DNA fragment encoding the P-450 type hydroxylase and ferredoxin genes was cloned from Pseudonocardia autotrophica IFO 12743 that can convert vitamin D$\_$3/ into its hydroxylated active forms. In order to isolate the P-450 gene cluster in this organism, we designed PCR primers on the basis of the regions of an oxygen binding site and a heme ligand pocket that are general characteristics of the P-450 hydroxylase. Sequencing analysis of the BamHI fragment revealed the presence of four complete and one incomplete ORFs, named PauA, PauB, PauC, and PauD, respectively. As a result of computer-based analyses, PauA and PauB have homology with enoyl-CoA hydratase from several organisms and the positive regulators belonging to the tetR family, respectively. PauC and PauD show similarity with SuaB/C proteins and ferredoxins, respectively, which are composed of P-450 monooxygenase systems for metabolizing two sulfonylurea herbicides in Streptomyces griseolus PauC shows the highest similarity with another CytP-450$\_$Sca2/ protein that is responsible for production of a specific HMG-CoA reductase inhibitor, pravastatin, in S. carbophilus. Cultures of Steptomyces lividans transformant, containing the P-450 gene cluster on the pWHM3 plasmid, was unable to convert vitamin D$\_$3/ to its hydroxylated forms.

Cloning and Functional Analysis of Gene Coding for S-Adenosyl-L-Methionine Synthetase from Streptomyces natalensis (Streptomyces natalensis로부터 S-adenosyl-L-methionine synthetase 유전자의 클로닝 및 기능분석)

  • Yoo, Dong-Min;Hwang, Yong-Il;Choi, Sun-Uk
    • Journal of Life Science
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    • 제21권1호
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    • pp.96-101
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    • 2011
  • S-Adenosyl-L-methionine synthtase (SAM-s) catalyzes the biosynthesis of SAM from ATP and L-methionine. SAM plays important roles in the primary and secondary metabolism of cells. A metK encoding a SAM-s was searched from Streptomyces natalensis producing natamycin, a predominantly a strong antifungal agent, inhibiting the growth of both yeasts and molds and preventing the formation of aflatoxin in filamentous fungi. To obtain the metK of S. natalensis, PCR using primers designed from the two highly conserved regions for metK genes of Streptomyces strains was carried out, and an intact 1.2-kb metK gene of S. natalensis was cloned by genomic Southern hybridization with PCR product as a probe. To identify the function of the cloned metK gene, it was inserted into pSET152ET for its high expression in the Streptomyces strain, and then introduced into S. lividans TK24 as a host by transconjugation using E. coli ET12567(pUZ8002). The high expression of metK in S. lividans TK24 induced actinorhodin production on R5 solid medium, and its amount in R4 liquid medium was 10-fold higher than that by exconjugant including only pSET152ET.

Distribution and Species Diversity of Wild Yeasts Isolated from Flowers in Korea (국내에 서식하는 꽃에서 분리한 야생 효모 분포 및 종 다양성)

  • Kim, Jeong-Seon;Lee, Miran;Kim, Jae Yoon;Heo, jun;Kwon, Soon-Wo;Yun, Bong Sik;Kim, Soo-Jin
    • The Korean Journal of Mycology
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    • 제48권4호
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    • pp.475-484
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    • 2020
  • Various indigenous yeasts were isolated and obtained from flowers in the Republic of Korea, and their distribution and species diversity were studied. Seventy-seven flowers were collected from 25 areas in Korea, and 502 yeast strains were isolated from these flowers. A total of 50 species were identified by comparing large subunit rDNA gene sequence homology with the type strains of yeasts. The analysis of yeast distribution showed that the dominant yeast species were Aureobasidium pullulans, A. leucospermi, and Filobasidium magnum in each region and flower samples. Except for the above three yeast species, no species of yeasts showed any meaningful distribution among the habitat regions and sources. In conclusion, 50 species of indigenous yeasts were obtained from flowers that can be used as industrial resources, and the data could be used for further research on yeast diversity and interactions between yeast and its host.

Endophytic Fungal Diversity Associated with the Roots of Coastal Sand-dune Plants in the Sindu-ri Coastal Sand Dune, Korea (신두리 해안사구에 자생하는 사구식물 내생진균의 다양성 분석)

  • You, Young-Hyun;Seo, Yeonggyo;Yoon, Hyeokjun;Kim, Hyun;Kim, Ye-Eun;Khalmuratova, Irina;Rim, Soon-Ok;Kim, Changmu;Kim, Jong-Guk
    • Microbiology and Biotechnology Letters
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    • 제41권3호
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    • pp.300-310
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    • 2013
  • The coastal sand-dune plants of eight species; Argusia sibirica, Calystegia soldanella, Elymus mollis, Lithospermum zollingeri, Raphanus sativus, Salsola collina, Zoysia macrostachya, and Zoysia sinica were collected from the Shindu-ri coastal sand dune. Ninety-eight endophytic fungal strains were isolated from the roots of these plants, analyzed, and identified by sequences in their internal transcribed spacers (ITS) at the ITS1, 5.8S, and ITS2 regions. The diversity of endophytic fungi isolated from coastal sand-dune plants was confirmed with various diversity indices. The fungal strains belonged to thirteen orders: Capnodiales (3.09%), Eurotiales (70.10%), Glomerellales (1.03%), Helotiales (3.09%), Hypocreales (9.28%), Mortierellales (2.06%), Onygenales (1.03%), Ophiostomatales (1.03%), Pleosporales (1.03%), Polyporales (1.03%), Russulales (1.03%), Saccharomycetales (2.06%), and Xylariales (1.03%). Of the endophytic fungal strains collected, Penicillium (59.18% in Eurotiales) and Fusarium (5.10% in Hypocreales) were the most abundant in coastal sand-dune plants. The endophytic fungal strains isolated from C. soldanella were more diverse compared to strains from the other coastal sand-dune plants.

Cellulose degrading basidiomycetes yeast isolated from the gut of grasshopper in Korea (한국의 메뚜기의 장에서 분리된 Cellulose를 분해하는 담자균 효모)

  • Kim, Ju-Young;Jang, Jun Hwee;Park, Ji-Hyun;Jung, Hee-Young;Park, Jong-Seok;Cho, Sung-Jin;Lee, Hoon Bok;Limtong, Savitree;Subramani, Gayathri;Sung, Gi-Ho;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • 제54권4호
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    • pp.362-368
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    • 2018
  • Grasshoppers play vital role in the digestion of photosynthetically fixed carbons. With the aid of intestinal microflora, the grasshopper can degrade leaves constituents such as cellulose and hemicellulose. The purpose of this study was to examine cellulolytic yeast isolates from the gut of grasshoppers collected in Gyeonggi Province, South Korea. Among the yeast isolates, ON2, ON17 (two strains), and ON6 (one strain) showed positive cellulolytic activity in the CMC-plate assay. The sequence analyses of D1/D2 domains of the large subunit rDNA gene and the internal transcribed spacer (ITS) regions revealed that the strains ON2 and ON17 were most closely related to Papiliotrema aspenensis CBS $13867^T$ (100%, sequence similarity in D1/D2 domains; 99.4% sequence similarity in ITS) and strain ON6 related to Saitozyma flava (100% in D1/D2 domains; 99.0% in ITS). All these three yeast strains are capable of degrading cellulose; therefore, the members of endosymbiotic yeasts may produce their own enzymes for carbohydrate degradation and convert mobilized sugar monomers to volatile fatty acids. Thus, the endosymbiotic yeast strains ON2, ON17 (represents the genus Papilioterma) and ON6 (Saitozyma) belonging to the family Tremellomycetes, are unreported strains in Korea.

Species identification and pathogenicity study of Colletotrichum isolates isolated from red-pepper and Chinese matrimony vine

  • Park, Eun-Sook;Lee, Bo-Heu;Min, Ji-Young;Cho, In-Joon;Kang, Beum-Kwan;Chung, Hae-Yeon;Yoo, Seung-Heon;Kim, Heung-Tae
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.131.1-131
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    • 2003
  • This study reports the identification of species of Colletotrichum strains originating from red-pepper and Chinese matrimony vine in Cheongyang. Ninteen isolates of red-pepper and 26 Coiletotrichum isolates of Chinese matrimony vine were compared with 5 isolates of strawberry representing C. gloeosporioides, by use of morphological and cultural criteria. Twenty three isolates among 26 isolates from Chinese matrimony vine were identified as C. acutatum, characterized by the low growth rates and the low sensitivity to carbendazim and diethofencarb. Also, all the isolates of red-pepper were identified as C. acutatum, showing the same characteristics as those of Chinese matrimony vine. Three and five isolates from Chinese matrimony vine and strawberry, respectively, were identified as C. gloeosporioides, characterized by the high growth rates and the high seneitivity to carbendazim and diethofencarb. There were differences in colony color and pathogenicity between Chinese matrimony vine isolates and red-pepper isolates of C. autatum. The isolates of C. acutatum from Chinese matrimony vine producing orange colored colonies with abundant spores showed the strong pathogenicity to Chinese matrimony vine, although they could not infect fruits of red-pepper by the wound inoculation. However, red-pepper isolates of C. acutatum producing gray colonies showed the strong pathogenicity to Chinese matrimony vine as well as red-pepper. Furthermore, comparative study on PCR amplification of ITS regions of rDNA was carried out using a number of Colletotrichum isolates. A species-specific primer could be used for the identification of C. acutatum from red-pepper and Chinese matrimony vine.

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Isolation of Lichen-forming Fungi from Hungarian Lichens and Their Antifungal Activity Against Fungal Pathogens of Hot Pepper Anthracnose

  • Jeon, Hae-Sook;Lokos, Laszlo;Han, Keon-Seon;Ryu, Jung-Ae;Kim, Jung-A;Koh, Young-Jin;Hur, Jae-Seoun
    • The Plant Pathology Journal
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    • 제25권1호
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    • pp.38-46
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    • 2009
  • Lichen-forming fungi (LEF) were isolated from 67 Hungarian lichen species from ascospores or thallus fragments. LFF were successfully isolated from 26 species with isolation rate of 38.8%. Of the total number of isolation from ascospores (27 species) and thallus fragments (40 species), 48% and 32.5% of the species were successfully isolated, respectively. Comparison of rDNA sequences of ITS regions between the isolated LFF and the original thallus confirmed that all the isolates originated from the thallus fragments were LEF. The following 14 species of LEF were newly isolated in this study; Acarospora cervina, Bacidia rubella, Cladonia pyxidata, Lasallia pustulata, Lecania hyaline, Lecanora argentata, Parmelina tiliacea, Parmotrema chinense, Physconia distorta, Protoparmeliopsis muralis, Ramalina pollinaria, Sarcogyne regularis, Umbilicaria hirsuta, Xanthoparmelia conspersa and X. stenophylla. Antifungal activity of the Hungarian LFF was evaluated against plant pathogenic fungi of Colletotrichum acutatum, C. coccodes and C. gloeosporioides, causal agent of anthracnose on hot pepper. Among the 26 isolates, 11 LFF showed more than 50% of inhibition rates of mycelial growth of at least one target pathogen. Especially, LFF of Evernia prunastri, Lecania hyalina and Lecanora argentata were remarkably effective in inhibition of mycelial growth of all the tested pathogens with antibiotic mode of action. On the other hands, five isolates of Cladonia furcata, Hypogymnia physodes, Lasallia pustulata, Ramalina fastigiata and Ramalina pollinaria exhibited fungal lytic activity against all the three pathogens. Among the tested fungal pathogens, C. coccodes seemed to be most sensitive to the LFF. The Hungarian LFF firstly isolated in this study can be served as novel bioresources to develop new biofungicides alternative to current fungicides to control hot pepper anthracnose pathogenic fungi.

Spatial Physicochemical and Metagenomic Analysis of Desert Environment

  • Sivakala, Kunjukrishnan Kamalakshi;Jose, Polpass Arul;Anandham, Rangasamy;Thinesh, Thangathurai;Jebakumar, Solomon Robinson David;Samaddar, Sandipan;Chatterjee, Poulami;Sivakumar, Natesan;Sa, Tongmin
    • Journal of Microbiology and Biotechnology
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    • 제28권9호
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    • pp.1517-1526
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    • 2018
  • Investigating bacterial diversity and its metabolic capabilities is crucial for interpreting the ecological patterns in a desert environment and assessing the presence of exploitable microbial resources. In this study, we evaluated the spatial heterogeneity of physicochemical parameters, soil bacterial diversity and metabolic adaptation at meter scale. Soil samples were collected from two quadrats of a desert (Thar Desert, India) with a hot, arid climate, very little rainfall and extreme temperatures. Analysis of physico-chemical parameters and subsequent variance analysis (p-values < 0.05) revealed that sulfate, potassium and magnesium ions were the most variable between the quadrats. Microbial diversity of the two quadrats was studied using Illumina bar-coded sequencing by targeting V3-V4 regions of 16S rDNA. As for the results, 702504 high-quality sequence reads, assigned to 173 operational taxonomic units (OTUs) at species level, were examined. The most abundant phyla in both quadrats were Actinobacteria (38.72%), Proteobacteria (32.94%), and Acidobacteria (9.24%). At genus level, Gaiella represented highest prevalence, followed by Streptomyces, Solirubrobacter, Aciditerrimonas, Geminicoccus, Geodermatophilus, Microvirga, and Rubrobacter. Between the quadrats, significant difference (p-values < 0.05) was found in the abundance of Aciditerrimonas, Geodermatophilus, Geminicoccus, Ilumatobacter, Marmoricola, Nakamurella, and Solirubrobacter. Metabolic functional mapping revealed diverse biological activities, and was significantly correlated with physicochemical parameters. The results revealed spatial variation of ions, microbial abundance and functional attributes in the studied quadrats, and patchy nature in local scale. Interestingly, abundance of the biotechnologically important phylum Actinobacteria, with large proposition of unclassified species in the desert, suggested that this arid environment is a promising site for bioprospection.