• Title/Summary/Keyword: rRNA sequence

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Characterization and Profiling of Liver microRNAs by RNA-sequencing in Cattle Divergently Selected for Residual Feed Intake

  • Al-Husseini, Wijdan;Chen, Yizhou;Gondro, Cedric;Herd, Robert M.;Gibson, John P.;Arthur, Paul F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1371-1382
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    • 2016
  • MicroRNAs (miRNAs) are short non-coding RNAs that post-transcriptionally regulate expression of mRNAs in many biological pathways. Liver plays an important role in the feed efficiency of animals and high and low efficient cattle demonstrated different gene expression profiles by microarray. Here we report comprehensive miRNAs profiles by next-gen deep sequencing in Angus cattle divergently selected for residual feed intake (RFI) and identify miRNAs related to feed efficiency in beef cattle. Two microRNA libraries were constructed from pooled RNA extracted from livers of low and high RFI cattle, and sequenced by Illumina genome analyser. In total, 23,628,103 high quality short sequence reads were obtained and more than half of these reads were matched to the bovine genome (UMD 3.1). We identified 305 known bovine miRNAs. Bta-miR-143, bta-miR-30, bta-miR-122, bta-miR-378, and bta-let-7 were the top five most abundant miRNAs families expressed in liver, representing more than 63% of expressed miRNAs. We also identified 52 homologous miRNAs and 10 novel putative bovine-specific miRNAs, based on precursor sequence and the secondary structure and utilizing the miRBase (v. 21). We compared the miRNAs profile between high and low RFI animals and ranked the most differentially expressed bovine known miRNAs. Bovine miR-143 was the most abundant miRNA in the bovine liver and comprised 20% of total expressed mapped miRNAs. The most highly expressed miRNA in liver of mice and humans, miR-122, was the third most abundant in our cattle liver samples. We also identified 10 putative novel bovine-specific miRNA candidates. Differentially expressed miRNAs between high and low RFI cattle were identified with 18 miRNAs being up-regulated and 7 other miRNAs down-regulated in low RFI cattle. Our study has identified comprehensive miRNAs expressed in bovine liver. Some of the expressed miRNAs are novel in cattle. The differentially expressed miRNAs between high and low RFI give some insights into liver miRNAs regulating physiological pathways underlying variation in this measure of feed efficiency in bovines.

Analysis of Microbial Diversity in Nuruk Using PCR-DGGE (PCR-DGGE를 이용한 누룩에서의 미생물 다양성 분석)

  • Kwon, Seung-Jik;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.1
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    • pp.110-116
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    • 2012
  • Nuruk plays a significant role in the flavor and quality of Takju and Yakju, which are produced through saccharification and alcohol fermentation by various microorganisms. In this study, we identified microbial strains isolated from a plate count and PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes, in order to characterize bacterial and fungal diversity in Sansung Nuruk. The numbers of bacteria and fungi in Nuruk were $1.5{\times}10^9$ CFU/g and $2.2{\tims}10^8$ CFU/g, respectively. The 16S rRNA gene sequence indicated that the predominant bacteria in the isolates and PCR-DGGE profile of Nuruk were Kocuria spp., Pantoea spp., Lactobacillus spp., Pediococcus spp., Weissella spp., Staphylococcus spp., endophytic bacterium, uncultured Gamma-proteobacteria, uncultured Cyanobacteria, and Actinobacteria. Dominant bacteria from the PCR-DGGE profile were Pediococcous pentosaceus and uncultured Cyanobacteria. The 28S rRNA gene sequence indicated the predominant fungi in the isolates and PCR-DGGE profile to be Trichomonascus spp. Pichia spp., Torulaspora spp., Wickerhamomyces spp., Sacharomycopsis spp., Lichtheimia spp., Mucor spp., Rhizopus spp. Aspergillus spp., and Cladosporium spp. Dominant fungi from the PCR-DGGE profile were Pichia kudriavzevii and Aspergillus oryzae. The PCR-DGGE technique was used for the first time in this study to assess a microbial community in Nuruk and proved to be an effective protocol for profiling microbial diversity.

Characterization of Korean Erwinia carotovora Strains from Potato and Chinese Cabbage

  • Seo, Sang-Tae;Koo, Jun-Hak;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.283-288
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    • 2004
  • Four Erwinia carotovora strains isolated from potatoes showing blackleg symptoms and rotted Chinese cabbage were analysed by biochemical tests and sequence analysis of 16S rDNA and 16S-23S rRNA intergenic spacer (IGS) regions, and the data were compared to related E. carotovora strains. Based on the results of the biochemical tests and sequence analysis, 2 of the 4 strains were identified as E. carotovora subsp. carotovora (Ecc), whereas the rest strains were distinct from Ecc. The last two strains, HCC3 and JEJU, were biochemically similar to E, carotovora subsp. atroseptica (Eca). However, the results of sequence analysis and Eca-specific PCR assays showed that the strains were distinct from Eca. On the basis of 16S rDNA sequence analysis, HCC3 and JEJU strains were placed in E. carotovora subsp. odorifera and E. carotovora subsp. wasabiae, respectively. The results of sequence analysis and specific PCR assay for Eca indicated that Asian Eca strains were distinct from European Eca strains, although they were phenotycally homogeneous.

Phylogenetic Relationships of Yeast-like Fungi Deduced from Partial Sequences of 18S Ribosomal RNA (18S 리보좀 RNA 부분 염기서열 분석에 의한 효모성 균류의 분자계통학적 연구)

  • Jung, Soo-Jin;Shin, Yong-Kook;Joo, Woo-Hong;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.23 no.4 s.75
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    • pp.310-317
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    • 1995
  • The evolutionary relationships for 43 species in the taxa of the basidiomycetous yeast and related yeast-like fungi have been studied from the 43 18S rRNA partial sequences. Fibulobasidium inconspicuum showed a relatively close relationship to Filobasidiella neoformans. It is obvious that Tremella foliacea and Ustilogo rabenhorstiana form a independently different cluster from basidiomycetous yeast and seem to be phylogenetically distant from basidiomycetous yeast and each other. The basidiomycetous yeasts are believed more closely associated with Tremella foliacea than with Ustilago rabenhorstiana. The phenotypic criteria such as carotenoid pigments, teliospores, ballistospores seems not to be phylogenetically useful indicators.

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Complete genome sequence of Flavobacteriaceae strain KCTC 52651 isolated from seawater recirculating aquaculture system (해수 순환여과양식시스템에서 분리된 Flavobacteriaceae 균주 KCTC 52651의 유전체 분석)

  • Kim, Young-Sam;Jeon, Young Jae;Kim, Kyoung-Ho
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.174-176
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    • 2019
  • A novel bacterium, designated strain RR4-38 (= KCTC 52651 = DSM 108068), belonging to the family Flavobacteriaceae was isolated from a biofilter in the seawater recirculating aquaculture system in South Korea. A single complete genome contig which is 3,182,272 bp with 41.9% G+C content was generated using PacBio RS II platform. The genome includes 2,829 protein-coding genes, 6 rRNA genes, 38 tRNA genes, 4 non-coding RNA genes, and 9 pseudogenes. The results will provide insights for understanding microbial activity in the seawater recirculating aquaculture system.

Development of Contig Assembly Program for Nucleotide Sequencing (염기서열 해독작업을 위한 핵산 단편 조립 프로그램의 개발)

  • 이동훈
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.121-127
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    • 1999
  • An effective computer program for assembling fragments in DNA sequencing has been developed. The program, called SeqEditor (Sequence Editor), is usable on the pcrsonal computer systems of MS-Widows which is the mosl popular operating system in Korea. It c'm recd several sequence file formats such as GenBak, FASTA, and ASCII. In the SeqEditor program, a dynamic programming algorihm is applied to compute the maximalscoring overlapping alignment between each pjlr of fragments. A novel feature of the program is that SeqEdilor implemnents interaclive operation with a graphical user interface. The performance lests of the prograln 011 fragmen1 data from 16s and 18s rDNA sequencing pi-ojects produced saiisIactory results. This program may be useful to a person who has work of time with large-scale DNA sequencing projects.

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Expression of a Cu-Zn Superoxide Dismutase Gene in Response to Stresses and Phytohormones in Rehmannia Glutinosa

  • Park, Myoung-Ryoul;Ryu, Sang-Soo;Yoo, Nam-Hee;Yu, Chang-Yeon;Yun, Song-Joong
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.270-275
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    • 2005
  • Superoxide dismutases (SOD) are metalloenzymes that convert $O_2^-\;to\;H_2O_2$. Rehmannia glutinosa is highly tolerant to paraquat-induced oxidative stress. The primary objective of this study was to characterize regulation of SOD gene expression in R. glutinosa in response to oxidative stresses and hormones. A full-length putative SOD clone (RgCu-ZnSOD1) was isolated from the leaf cDNA library of R. glutinosa using an expressed sequence tag clone as a probe. RgCu-ZnSOD1 cDNA is 777 bp in length and contains an open reading frame for a polypeptide consisted of 152 amino acid residues. The deduced amino acid sequence of the clone shows highest sequence similarity to the cytosolic Cu-ZnSODs. The two to three major bands with several minor ones on the Southern blots indicate that RgCu-ZnSOD1 is a member of a small multi-gene family. RgCuZnSOD1 mRNA was constitutively expressed in the leaf, flower and root. The expression of RgCu-ZnSOD1 mRNA was increased about 20% by wounding and paraquat, but decreased over 50% by ethylene and $GA_3$. This result indicates that the RgCu-ZnSOD1 expression is regulated differentially by different stresses and phytohormones at the transcription level. The RgCu-ZnSOD1 sequence and information on its regulation will be useful in investigating the role of SOD in the paraquat tolerance of R. glutinosa.

The complete mitochondrial genome sequence of the indigenous I pig (Sus scrofa) in Vietnam

  • Nguyen, Hieu Duc;Bui, Tuan Anh;Nguyen, Phuong Thanh;Kim, Oanh Thi Phuong;Vo, Thuy Thi Bich
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.930-937
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    • 2017
  • Objective: The I pig is a long nurtured longstanding breed in Vietnam, and contains excellent indigenous genetic resources. However, after 1970s, I pig breeds have become a small population because of decreasing farming areas and increasing pressure from foreign breeds with a high growth rate. Thus, there is now the risk of the disappearance of the I pigs breed. The aim of this study was to focus on classifying and identifying the I pig genetic origin and supplying molecular makers for conservation activities. Methods: This study sequenced the complete mitochondrial genome and used the sequencing result to analyze the phylogenetic relationship of I pig with Asian and European domestic pigs and wild boars. The full sequence was annotated and predicted the secondary tRNA. Results: The total length of I pig mitochondrial genome (accession number KX094894) was 16,731 base pairs, comprised two rRNA (12S and 16S), 22 tRNA and 13 mRNA genes. The annotation structures were not different from other pig breeds. Some component indexes as AT content, GC, and AT skew were counted, in which AT content (60.09%) was smaller than other pigs. We built the phylogenetic trees from full sequence and D loop sequence using Bayesian method. The result showed that I pig, Banna mini, wild boar (WB) Vietnam and WB Hainan or WB Korea, WB Japan were a cluster. They were a group within the Asian clade distinct from Chinese pigs and other Asian breeds in both phylogenetic trees (0.0004 and 0.0057, respectively). Conclusion: These results were similar to previous phylogenic study in Vietnamese pig and showed the genetic distinctness of I pig with other Asian domestic pigs.

Differentiation of three scuticociliatosis causing species in olive flounder (Paralichthys olivaceus) by multiplex PCR

  • Kim, Sung Mi;Lee, Eun Hye;Kim, Ki Hong
    • Journal of fish pathology
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    • v.17 no.2
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    • pp.145-149
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    • 2004
  • The definitive identification of ciliate species by morphological characteristics relies on time-consuming and laborious staining techniques. Therefore, in this study, we discriminated 3 scuticociliatosis causing species - Pseudocohnilembus persalinus, Uronema marinum and Philasterides dicentrarchi - in cultured olive flounder by multiplex PCR. The multiplex PCR based on the species-specific amplification of small subunit ribosomal RNA (SS rRNA) gene sequence enabled us to distinguish the 3 scuticociliate species in a simple and rapid manner, even in the sample containing the three species simultaneously. These data suggest that the multiplex PCR strategy would make it possible to avoid the cumbersome and time-consuming procedures of morphological analysis for the definitive identification of scuticociliates.

Phylogenetic Diversity and Community Analysis of Marine Bacteria Associated with Ulva pertusa (구멍갈파래(Ulva pertusa)에 서식하는 해양세균의 계통학적 다양성 및 군집구조 분석)

  • Choi, Ha-Ri;Park, So-Hyun;Kim, Dong-Hwi;Kim, Ji-Young;Heo, Moon-Soo
    • Journal of Life Science
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    • v.26 no.7
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    • pp.819-825
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    • 2016
  • The present study was done to assess the diversity of the bacterial community associated with Ulva pertusa collected from Jeju Island using Restriction Fragment Length Polymorphism (RFLP) marker analysis. For RFLP analysis, a total of 145 bacterial strains associated with Ulva pertusa were screened and cultivated using Marine agar and R2A agar. The PCR amplicons of the 16S rRNA gene from all the isolated strains were digested with HaeIII and RsaI restriction enzymes and then classified into different groups according to their restriction patterns. Strains selected based on the RFLP patterns showed more than 91% 16S rRNA gene sequence similarity when compared with known bacterial species, which include 4 phyla - proteobacteria (alpha-proteobacteria, beta-proteobacteria, gamma-proteobacteria - 63%), firmicutes (11%), actinobacteria (4%), bacteroidetes (22%)–as well as 7 classes (actinobacteria, flavobacteriia, cytophagia, bacilli, α-proteobacteria, γ-proteobacteria, β-proteobacteria), 13 orders, 18 families, and 27 genera. These results confirmed a wide diversity of bacterial communities as contrasted with other regions. The newly isolated 10 strains, which show 16S rRNA sequence similarity of <97% compared to previously identified bacteria, could be noble species. Further experiments, such as morphological, physiological, and biochemical classification, are necessary to confirm the novelty of the newly isolated 10 strains.