• Title/Summary/Keyword: RNA sequencing

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Temporal expression profiling of long noncoding RNA and mRNA in the peripheral blood during porcine development

  • Gu, Yiren;Zhou, Rui;Jin, Long;Tao, Xuan;Zhong, Zhijun;Yang, Xuemei;Liang, Yan;Yang, Yuekui;Wang, Yan;Chen, Xiaohui;Gong, Jianjun;He, Zhiping;Li, Mingzhou;Lv, Xuebin
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.5
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    • pp.836-847
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    • 2020
  • Objective: We investigated the temporal expression profiles of long noncoding RNA (lncRNA) and mRNA in the peripheral blood of pigs during development and identified the lncRNAs that are related to the blood-based immune system. Methods: Peripheral blood samples were obtained from the pigs at 0, 7, 28, and 180 days and 2 years of age. RNA sequencing was performed to survey the lncRNA and mRNA transcriptomes in the samples. Short time-series expression miner (STEM) was used to show temporal expression patterns in the mRNAs and lncRNAs. Gene ontology and Kyoto encyclopedia of genes and genomes analyses were performed to assess the genes' biological relevance. To predict the functions of the identified lncRNAs, we extracted mRNAs that were nearby loci and highly correlated with the lncRNAs. Results: In total of 5,946 lncRNA and 12,354 mRNA transcripts were identified among the samples. STEM showed that most lncRNAs and mRNAs had similar temporal expression patterns during development, indicating the expressional correlation and functional relatedness between them. The five stages were divided into two classes: the suckling period and the late developmental stage. Most genes were expressed at low level during the suckling period, but at higher level during the late stages. Expression of several T-cell-related genes increased continuously during the suckling period, indicating that these genes are crucial for establishing the adaptive immune system in piglets at this stage. Notably, lncRNA TCONS-00086451 may promote blood-based immune system development by upregulating nuclear factor of activated T-cells cytoplasmic 2 expression. Conclusion: This study provides a catalog of porcine peripheral blood-related lncRNAs and mRNAs and reveals the characteristics and temporal expression profiles of these lncRNAs and mRNAs during peripheral blood development from the newborn to adult stages in pigs.

Site-Specific Mutagenesis on the 32-T and 39-T of E. coli $tRNA^{phe}$ Gene (E. coli $tRNA^{phe}$ 유전자의 32-T와 39-T 염기의 부의 특이적 돌연변이)

  • 김익영;이세영
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.176-180
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    • 1989
  • There are three pseudourdine ($Psi$)bases in the E. coli $tRNA^{phe}$ In order to study the function of the pseudouridine bases in the $tRNA^{phe}$, changes of bases $tRNA^{phe}$ gene to other bases were undertaken by the site-specific mutagenesis. Site-specific mutagenesis of T in the pheW gene, a $tRNA^{phe}$ gene of E. coli, corresponding to the baseat the No.32 position to C and also T corresponding to the base at the No.39 position to C were performed using Kunkel's uracil-containing template method. Identification of mutants were undertaken by the KNA sequencing techniques of the mutated pheW genes and activities of the mutated pheW genes complementing to E. coli NP37 mutant($pheS^{-ts}$) using the recombinant plasmid containing the mutated genes. Neither NP37 harboring pheW gene mutated at No.32 position nor NP37 harboring pheW gene mutated at No.39 position can be grown at non-permissive temperature. The result means that both mutated pheW genes can not complement to E. coli NP37, and that the pseudouridine bases are essential to the activity of the E. coli $tRNA^{phe}$ in vivo.

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Characterization of beer-spoilage microorganism and its rapid detection by specific PCR primer (맥주오염미생물의 동정과 specific PCR primer의한 신속한 검출 방법)

  • Lee, Taek-In;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.28 no.A
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    • pp.141-147
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    • 2008
  • Several contaminated bacteria such as Lactobacillus brevis and Pediococcus damnosus in beer production cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. Recently, two contaminated strains were isolated from vessel of beer production and identified as Lactobacillus species by API kit identificaton as well as 16S-23S ITS sequencing analyses. Two isolated strains were named as Lactobacillus sp. HLA1 and Lactobacillus HLB2, respectively. A polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Lactobacillus sp.. Two sets of primer pairs (HLA1-F/HLA1-R and HLB2-F/HLB2-R) were designed for the amplification of a 1576 base pair (bp) fragment of the HLA1 16S-23S rRNA gene and 1888 bp fragement of the HLB2 16S-23S rRNA. Amplified PCR products were highly specific to detect corresponding bacteria when other contaminated strains were used as PCR templates. However, detection of both strains were limited when $100{\mu}{\ell}$ of cultured samples were mixed with $100m{\ell}$ of beer sample in arbitrary manner. The sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

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Malassezia Dermatitis in a South American Sea Lion (Otaria byronia) (오타리아 물개(South American Sea Lion, Otaria byronia)에서 발생한 Malassezia 피부염)

  • Ahn, Miji;Bae, Seulgi;Oh, Taeho
    • Journal of Veterinary Clinics
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    • v.31 no.2
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    • pp.156-158
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    • 2014
  • Malassezia spp. was isolated from the skin lesion of a South American Sea Lion (Otaria byronia) in South Korea. Colonies were cream to yellowish with smooth margin on Sabouraud dextrose agar without lipid supplements. Yeast cells were ovoid to cylindrical in shape and budding daughter cells from broad base. We conducted 26s rRNA sequencing to specify the strain of the yeast and found out this isolate highly matched with Malassezia pachydermatis isolated from canine otitis. The pulse therapy of oral itraconazole was very effective in this case. M. pachydermatis has a wide range of host animals but this is the first report in a sea lion in South Korea.

Detection, isolation, and characterization of the cucumber mosaic virus in Pseudostellaria heterophylla from Korea

  • Lee, Da Hyun;Kim, Jinki;Han, Jun Soo;Lee, Jae-Hyeon;Lee, ByulHaNa;Park, Chung Youl
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.150-156
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    • 2020
  • Weeds play an important role in the survival of viruses and are potential inoculum sources of viral diseases for crop plants. In this study, specimens of Pseudostellaria heterophylla exhibiting symptoms of the cucumber mosaic virus (CMV) were collected in Bonghwa, Korea. The characteristics of the disease were described and leaf RNA was extracted and sequenced to identify the virus. Three CMV contigs were obtained and PCR was performed using specific primer pairs. RNA from positive samples exhibiting CMV leaf symptoms was amplified to determine the coat protein. A sequence comparison of the coat protein gene from the CMV BH isolate shared the highest nucleotide identity (99.2%) with the CMV ZM isolate. Phylogenetic analysis showed that CMV-BH belonged to subgroup IA and that the most closely-related isolate was CMV-ZM. All test plants used for the biological assay were successfully infected with CMV and exhibited CMV disease symptoms such as blistering, mosaic, and vein yellowing. To our knowledge, this is the first report of CMV infection in P. heterophylla from Korea.

A Case of Furuncular Myiasis Due to Cordylobia anthropophaga in a Korean Traveler Returning from Uganda

  • Song, Su-Min;Kim, Shin-Woo;Goo, Youn-Kyoung;Hong, Yeonchul;Ock, Meesun;Cha, Hee-Jae;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.55 no.3
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    • pp.327-331
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    • 2017
  • A fly larva was recovered from a boil-like lesion on the left leg of a 33-year-old male on 21 November 2016. He has worked in an endemic area of myiasis, Uganda, for 8 months and returned to Korea on 11 November 2016. The larva was identified as Cordylobia anthropophaga by morphological features, including the body shape, size, anterior end, posterior spiracles, and pattern of spines on the body. Subsequent 28S rRNA gene sequencing showed 99.9% similarity (916/917 bp) with the partial 28S rRNA gene of C. anthropophaga. This is the first imported case of furuncular myiasis caused by C. anthropophaga in a Korean overseas traveler.

Effect of random Shine-Dalarno sequence on the expression of Bovine Growth Hormone Gene in Escherichia coli (대장균에서 무작위 샤인-달가노 서열이 소성장호르몬 유전자 발현에 미치는 영향)

  • 나경수;나경수;백형석;이용세
    • Journal of Life Science
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    • v.10 no.4
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    • pp.422-430
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    • 2000
  • In order to search for the effects of Shine-Dalgarno (SD) sequence and nucleotide sequence of spacer region (SD-ATG) on bGH expression, oligonucleotides containing random SD sequences and a spacer region were chemically synthesized. The distance between SD region and initiation codon (ATG) was fixed to 9 nucleotides in length. The expression vectors have been constructed using pT7-1 vector containing a T7 promoter. Positive clones were screened with colony hybridization and named pT7A or pT7B plasmid series. The selected clones were confirmed by DNA sequencing and finally, 19 clones having various SD combinations were obtained. When bovine growth hormone was induced by IPTG in E. coli BL21(DE3), all cells harboring these plasmids produced a detectable level of bGH in western blot analysis. However, various SD sequences did not affect on bGH expression, indicating that the sequences of SD and the spacer region did not sufficiently destabilize mRNA secondary structure of bGH gene. Therefore, these results indicate that the disruption of mRNA secondary structure might be a major factor for regulating bGH expression in the translational initiation process.

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Cloning and Sequencing of Resistance Determinants to Aminoglycoside Antibiotics from Sterptoalloteichus hindustanus ATCC 31219 (Streptoalloteichus hindustanus ATCC 31219로부터 아미노글라이코사이드계 항생제에 내성을 지정하는 유전자의 클로닝 및 염기서열 결정)

  • Kim, Jong-Woo;Han, Jae-Jin;Choi, Young-Nae;Eom, Joon-Ho;Yoon, Sung-Joon;Hyun, Chang-Gu;Suh, Joo-Won
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.384-389
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    • 1995
  • Streptoalloteichus hindustanus ATCC 31219, a nebramycin complex producer, is similar to Streptomyeces tenebrarius in a viewpoint of resistance to a wide range of aminoglycoside antibiotics. S. tenebrarius has resistance mechanisms of 16s rRNA methylation and aminogycoside modification. However, it is not known whether resistance mechanisms of Stall. hindustanus are the same as in S. tenebrarius. Therefore, we have tried to isolate resistance determinants from Stall. hindustanus. Two different types of aminoglycoside resistance determinants were isolated from Stall. hindustanus and expressed in Streptomyces lividans TK24. The apramycin resistance gene (amr) and the tobramycin resistance gene (tmr) isolated from Stall. hindustanus showed broad resistance spectrum against a dozen of aminoglycoside antibiotics. The complete nucleotide sequences of apramycin resistance gene (amr) were determined. The deduced amino acid sequence of the amr gene of Stall hindustanus ATCC 31219 showed extensive sequence homology to the 16s rRNA methylase gene (kamB) of S. tenebrarius.

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Beyond gene expression level: How are Bayesian methods doing a great job in quantification of isoform diversity and allelic imbalance?

  • Oh, Sunghee;Kim, Chul Soo
    • Journal of the Korean Data and Information Science Society
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    • v.27 no.1
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    • pp.225-243
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    • 2016
  • Thanks to recent advance of next generation sequencing techniques, RNA-seq enabled to have an unprecedented opportunity to identify transcript variants with isoform diversity and allelic imbalance (Anders et al., 2012) by different transcriptional rates. To date, it is well known that those features might be associated with the aberrant patterns of disease complexity such as tissue (Anders and Huber, 2010; Anders et al., 2012; Nariai et al., 2014) specific differential expression at isoform levels or tissue specific allelic imbalance in mal-functionality of disease processes, etc. Nevertheless, the knowledge of post-transcriptional modification and AI in transcriptomic and genomic areas has been little known in the traditional platforms due to the limitation of technology and insufficient resolution. We here stress the potential of isoform variability and allelic specific expression that are relevant to the abnormality of disease mechanisms in transcriptional genetic regulatory networks. In addition, we systematically review how robust Bayesian approaches in RNA-seq have been developed and utilized in this regard in the field.