• Title/Summary/Keyword: RNA sequencing

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Molecular characteristics of diverse dsRNAs in edible fungi (버섯에 존재하는 다양한 dsRNA의 분자생물학적 연구)

  • Park, Yunjung;Shin, Pyung-Gyun;Jang, Kab-Yeul;Kong, Won-Sik;Cheong, Jong-Chun;Yoo, Young-Bok
    • Journal of Mushroom
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    • v.8 no.2
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    • pp.41-50
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    • 2010
  • Mycoviruses have been found in many fungal species including mushrooms. Double-stranded (ds) RNA genomes were common type in mycoviruses, but single-stranded (ss) RNA mycoviruses were also reported in some fungal species. Sequencing analysis using cDNA cloning experiments revealed that mycoviruses can be classified into several different virus families such as Totiviridae, Hypoviridae, Partitiviridae and Barnaviridae etc. Because the nucleotide sequence data that are available in these days are very limited in a number of mycoviruses, the existence of more diverse viral groups in fungi are currently expected. In this review, we selected four different fungal groups, which were considered as the model systems for mycovirus related studies in both plant pathogenic fungi and edible mushroom species, and discussed about their molecular characteristics of diverse mycoviruses. The plant pathogenic fungi introduced here were Cryphonectria parasitica and Helminthosporium victoriae and the edible mushroom species were Agaricus bisporus and Pleurotus ostreatus.

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Eight unrecorded bacterial species isolated from soil and marine sediment in Korea

  • Kim, Minji;Lee, Ki-Eun;Cha, In-Tae;Lee, Byoung-Hee;Park, Soo-Je
    • Journal of Species Research
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    • v.9 no.4
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    • pp.339-345
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    • 2020
  • The Earth contains billions of microbial species, although the vast majority cannot be cultured in laboratories and are thus considered unidentified and uncharacterized. Extremophiles are microorganisms that thrive in extreme conditions, including temperature, salinity, and pH. Extremophilic microorganisms have provided important insights for biological, metabolic, and evolutionary studies. Between 2017 and 2019, as part of a comprehensive investigation to identify bacterial species in Korea, eight bacterial strains were isolated from marine and non-marine environments in Jeju Island. These strains were cultured under extreme salinity or pH conditions. Phylogenetic analysis using 16S ribosomal RNA(rRNA) gene sequencing indicated that all eight strains belonged to the phyla Gammaproteobacteria, Bacilli, and Alphaproteobacteria. Based on their high 16S rRNA gene sequence similarities(>98.7%) and the formation of strong monophyletic clades with their closest related species, all isolated strains were considered as an unrecorded strain, previously unidentified species. Gram stain reaction, culture conditions, colony and cell morphology, biochemical characteristics, isolation source, and National Institute of Biological Resources(NIBR) IDs are described in this article. The characterization of these unrecorded strains provides information on microorganisms living in Korea.

Cancer-Specific Induction of Adenoviral E1A Expression by Group I Intron-Based Trans-Splicing Ribozyme

  • Won, You-Sub;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.431-435
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    • 2012
  • In this study, we describe a novel approach to achieve replicative selectivity of conditionally replicative adenovirus that is based upon trans-splicing ribozyme-mediated replacement of cancer-specific RNAs. We developed a specific ribozyme that can reprogram human telomerase reverse transcriptase (hTERT) RNA to induce adenoviral E1A gene expression selectively in cancer cells that express the RNA. Western blot analysis showed that the ribozyme highly selectively triggered E1A expression in hTERT-expressing cancer cells. RT-PCR and sequencing analysis indicated that the ribozyme-mediated E1A induction was caused via a high fidelity trans-splicing reaction with the targeted residue in the hTERT-expressing cells. Moreover, reporter activity under the control of an E1A-dependent E3 promoter was highly transactivated in hTERT-expressing cancer cells. Therefore, adenovirus containing the hTERT RNA-targeting trans-splicing ribozyme would be a promising anticancer agent through selective replication in cancer cells and thus specific destruction of the infected cells.

Bioinformatics Approach to Direct Target Prediction for RNAi Function and Non-specific Cosuppression in Caenorhabditis elegans (생물정보학적 접근을 통한 Caenorhabditis elegans 모델시스템의 생체내 RNAi 기능예측 및 비특이적 공동발현억제 현상 분석)

  • Kim, Tae-Ho;Kim, Eui-Yong;Joo, Hyun
    • KSBB Journal
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    • v.26 no.2
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    • pp.131-138
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    • 2011
  • Some computational approaches are needed for clarifying RNAi sequences, because it takes much time and endeavor that almost of RNAi sequences are verified by experimental data. Incorrectness of RNAi mechanism and other unaware factors in organism system are frequently faced with questions regarding potential use of RNAi as therapeutic applications. Our massive parallelized pair alignment scoring between dsRNA in Genebank and expressed sequence tags (ESTs) in Caenorhabditis elegans Genome Sequencing Projects revealed that this provides a useful tool for the prediction of RNAi induced cosuppression details for practical use. This pair alignment scoring method using high performance computing exhibited some possibility that numerous unwanted gene silencing and cosuppression exist even at high matching scores each other. The classifying the relative higher matching score of them based on GO (Gene Ontology) system could present mapping dsRNA of C. elegans and functional roles in an applied system. Our prediction also exhibited that more than 78% of the predicted co-suppressible genes are located in the ribosomal spot of C. elegans.

Elucidating molecular mechanisms of acquired resistance to BRAF inhibitors in melanoma using a microfluidic device and deep sequencing

  • Han, Jiyeon;Jung, Yeonjoo;Jun, Yukyung;Park, Sungsu;Lee, Sanghyuk
    • Genomics & Informatics
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    • v.19 no.1
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    • pp.2.1-2.10
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    • 2021
  • BRAF inhibitors (e.g., vemurafenib) are widely used to treat metastatic melanoma with the BRAF V600E mutation. The initial response is often dramatic, but treatment resistance leads to disease progression in the majority of cases. Although secondary mutations in the mitogen-activated protein kinase signaling pathway are known to be responsible for this phenomenon, the molecular mechanisms governing acquired resistance are not known in more than half of patients. Here we report a genome- and transcriptome-wide study investigating the molecular mechanisms of acquired resistance to BRAF inhibitors. A microfluidic chip with a concentration gradient of vemurafenib was utilized to rapidly obtain therapy-resistant clones from two melanoma cell lines with the BRAF V600E mutation (A375 and SK-MEL-28). Exome and transcriptome data were produced from 13 resistant clones and analyzed to identify secondary mutations and gene expression changes. Various mechanisms, including phenotype switching and metabolic reprogramming, have been determined to contribute to resistance development differently for each clone. The roles of microphthalmia-associated transcription factor, the master transcription factor in melanocyte differentiation/dedifferentiation, were highlighted in terms of phenotype switching. Our study provides an omics-based comprehensive overview of the molecular mechanisms governing acquired resistance to BRAF inhibitor therapy.

The Biological Functions of Plant Long Noncoding RNAs (식물의 긴비암호화 RNA들의 생물학적 기능)

  • Kim, Jee Hye;Heo, Jae Bok
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1097-1104
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    • 2016
  • With the development of next generation sequencing (NGS), large numbers of transcriptional molecules have been discovered. Most transcripts are non -coding RNAs (ncRNAs). Among them, long non-coding RNAs (lncRNAs) with more than 200 nucleotides represent functional RNA molecule that will not be translated into protein. In plants, lncRNAs are transcribed by RNA polymerase II (Pol II) or Pol III, Pol VI and Pol V. After transcription of these lncRNAs, more RNA processing mechanisms such as splicing and polyadenylation occurs. The expression of plant lncRNAs is very low and is tissue specific. However, these lncRNAs are strongly induced by specific external stimuli. Because different external stimuli including environmental stresses induce a large number of plant lncRNAs, these lncRNAs have been gradually considered as new regulatory factors of various biological and development processes such as epigenetic repression, chromatin modification, target mimicry, photomorphogenesis, protein relocalization, environmental stress response, pathogen infection in plants. Moreover, some lncRNAs act as precursor of short RNAs. Although a large number of lncRNAs have been predicted and identified in plants, our current understanding of the biological function of these lncRNAs is still limited and their detailed regulatory mechanisms should be elucidated continuously. Here, we reviewed the biogenesis and regulation mechanisms of lncRNAs and summarized the molecular functions unraveled in plants.

Sequence Verification of Synthetic Oligonucleotides by Exonuclease Digestion and Matrix Assisted Laser Desorption Ionization Mass Spectrometry

  • Kim, Jin-Sung;Jang, Jung-Suk;Choi, Jong-Soon;Chang, Yoon-Seok
    • BMB Reports
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    • v.29 no.2
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    • pp.122-126
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    • 1996
  • A series of oligonucleotides were synthesized by automatic DNA synthesizer. The purity of crude products was checked and their molecular weights determined by matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) with an accuracy of better than 0.05% deviation even without using an internal standard. This mass determining technology in combination with partial digestion of oligonucleotides by 5'- and 3'-exonuclease provides a straightforward and simple method to obtain sequence information of oligonucleotides. The extension of this technology to the sequencing of modified oligonucleotides and genomic DNA and RNA might become possible.

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A Case of Plasmodium ovale Malaria Imported from West Africa

  • Kang, Yunjung;Yang, Jinyoung
    • Parasites, Hosts and Diseases
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    • v.51 no.2
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    • pp.213-218
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    • 2013
  • Malaria is a parasitic infection caused by Plasmodium species. Most of the imported malaria in Korea are due to Plasmodium vivax and Plasmodium falciparum, and Plasmodium ovale infections are very rare. Here, we report a case of a 24-year-old American woman who acquired P. ovale while staying in Ghana, West Africa for 5 months in 2010. The patient was diagnosed with P. ovale malaria based on a Wright-Giemsa stained peripheral blood smear, Plasmodium genus-specific real-time PCR, Plasmodium species-specific nested PCR, and sequencing targeting 18S rRNA gene. The strain identified had a very long incubation period of 19-24 months. Blood donors who have malaria with a very long incubation period could be a potential danger for propagating malaria. Therefore, we should identify imported P. ovale infections not only by morphological findings but also by molecular methods for preventing propagation and appropriate treatment.

Sequencing of Enzymatically Amplified DNA and Its Application to 16S Like Ribosomal Gene Amplification and Sequencing (효소적으로 증폭된 DNA의 염기배열법과 16S like 리보좀 유전자의 증폭 및 염기배열결정에의 응용)

  • 이재동;주우홍
    • Journal of Life Science
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    • v.2 no.2
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    • pp.108-119
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    • 1992
  • 근년에 개발된 효소적인 DNA증폭법을 이용하면 일차구조상의 단편적인 정보만 알면 단 수시간내 해석에 필요한 양의 DNA가 증폭되어 cDNA의 염기배열결정의 신속화, 간편화가 가능하게 되었다. 그러므로 유전자증폭법으로써 PCR법에 관해 기술한다. 그리고 리보좀 RNA는 분자시계로서 생물의 계통을 논하는 데에 있어서는 최적의 조건을 갖춘 고분자화합물이다. 이에 PCR법을 이용한 16S like 리보좀DNA의 증폭법을 다루고, PCR증폭산물의 염기배열결정법에 대해 서술한다. 또한 인위적인 leading error 등을 배제하고 신속한 자동해독과 시간적인 절약이 자동 DNA sequencer의 개발과 시판으로 가능하게 되어 cDNA의 형광색소표식 염기배열결정법에 대해서도 서술한다.

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Molecular cloning and nucleotide sequencing of bovine viral diarrhea virus gp53 antigenic region (소 바이러스성 설사병 바이러스 gp53 항원부위 유전자의 재조합 및 염기서열 연구)

  • Yeo, Sang-geon;Cho, HJ;Masri, SA
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.287-295
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    • 1995
  • Molecular cloning and nucleotide sequencing were undertaken for the RNA genome of gp53 antigenic region in cytopathic Singer strain of bovine viral diarrhea virus. The cloned cDNA was 939 nucleotides in length having a base composition of 31.0% A, 19.6% C, 25.5% G and 24.0% T. The sequence was corresponded to approximately 77.8%(817 bases) of predicted gp53 region and 122 bases after 3'end of gp53 region in the Singer strain when compared with NADL strain of known sequence. A single open reading frame was found in the sequence of 2nd frame and was deduced as encoding 312 amino acids.

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