• Title/Summary/Keyword: Transcription Method

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Regulatory Network of MicroRNAs, Host Genes, Target Genes and Transcription Factors in Human Esophageal Squamous Cell Carcinoma

  • Wang, Tian-Yan;Xu, Zhi-Wen;Wang, Kun-Hao;Wang, Ning
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3677-3683
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    • 2015
  • Abnormally expressed microRNAs (miRNAs) and genes have been found to play key roles in esophageal squamous cell carcinoma (ESCC), but little is known about the underlying mechanisms. The aim of this paper was to assess inter-relationships and the regulatory mechanisms of ESCC through a network-based approach. We built three regulatory networks: an abnormally expressed network, a related network and a global network. Unlike previous examples, containing information only on genes or miRNAs, the prime focus was on relationships. It is worth noting that abnormally expressed network emerged as a fault map of ESCC. Theoretically, ESCC might be treated and prevented by correcting the included errors. In addition, the predicted transcription factors (TFs) obtained by the P-match method also warrant further study. Our results may further guide gene therapy researchers in the study of ESCC.

G-Networks Based Two Layer Stochastic Modeling of Gene Regulatory Networks with Post-Translational Processes

  • Kim, Ha-Seong;Gelenbe, Erol
    • Interdisciplinary Bio Central
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    • v.3 no.2
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    • pp.8.1-8.6
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    • 2011
  • Background: Thanks to the development of the mathematical/statistical reverse engineering and the high-throughput measuring biotechnology, lots of biologically meaningful genegene interaction networks have been revealed. Steady-state analysis of these systems provides an important clue to understand and to predict the systematic behaviours of the biological system. However, modeling such a complex and large-scale system is one of the challenging difficulties in systems biology. Results: We introduce a new stochastic modeling approach that can describe gene regulatory mechanisms by dividing two (DNA and protein) layers. Simple queuing system is employed to explain the DNA layer and the protein layer is modeled using G-networks which enable us to account for the post-translational protein interactions. Our method is applied to a transcription repression system and an active protein degradation system. The steady-state results suggest that the active protein degradation system is more sensitive but the transcription repression system might be more reliable than the transcription repression system. Conclusions: Our two layer stochastic model successfully describes the long-run behaviour of gene regulatory networks which consist of various mRNA/protein processes. The analytic solution of the G-networks enables us to extend our model to a large-scale system. A more reliable modeling approach could be achieved by cooperating with a real experimental study in synthetic biology.

Rapid and Specific Detection of Apple stem grooving virus by Reverse Transcription-recombinase Polymerase Amplification

  • Kim, Nam-Yeon;Oh, Jonghee;Lee, Su-Heon;Kim, Hongsup;Moon, Jae Sun;Jeong, Rae-Dong
    • The Plant Pathology Journal
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    • v.34 no.6
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    • pp.575-579
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    • 2018
  • Apple stem grooving virus (ASGV) is considered to cause the most economically important viral disease in pears in Korea. The current PCR-based methods used to diagnose ASGV are time-consuming in terms of target detection. In this study, a novel assay for specific ASGV detection that is based on reverse transcription-recombinase polymerase amplification is described. This assay has been shown to be reproducible and able to detect as little as $4.7ng/{\mu}l$ of purified RNA obtained from an ASGV-infected plant. The major advantage of this assay is that the reaction for the target virus is completed in 1 min, and amplification only requires an incubation temperature of $42^{\circ}C$. This assay is a promising alternative method for pear breeding programs or virus-free certification laboratories.

Detection of Marine Birnavirus (MBV) from Rockfish Sebastes schlegeli Using Reverse Transcription and Nested PCR

  • Joh, Seong-Joon;Kim, Doo-Won;Kim, Jeong-Ho;Heo, Gang-Joon
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.260-264
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    • 2000
  • Reverse transcription (RT)-PCR and nested PCR methods (2-step PCR) were tested for their ability to detect marine birnavirus (MBV) in cultured rockfish, Sebastes schlegeli. One set of primers for RT-PCR was designed, based on a gene of infectious pancreatic necrosis virus (IPNV), and another set of primers for nested PCR was designed based on the VP2/NS junction region of MBV. This 2-step PCR method was specific for MBV and sensitivity was heightened when nested PCR was combined to RT-PCR. This 2-step PCR method was useful for detecting MBV not only in diseased fish, but also in asymptomatic fish. These results indicate that this 2-step PCR method is useful for detecting MBV in rockfish.

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RT-RPA Assay Combined with a Lateral Flow Strip to Detect Soybean Mosaic Virus

  • Bong Geun Oh;Ju-Yeon Yoon;Ho-Jong Ju
    • The Plant Pathology Journal
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    • v.40 no.4
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    • pp.337-345
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    • 2024
  • Soybean (Glycine max L.) is one of the most widely planted and used legumes in the world, being used for food, animal feed products, and industrial production. The soybean mosaic virus (SMV) is the most prevalent virus infecting soybean plants. This study developed a diagnostic method for the rapid and sensitive detection of SMV using a reverse transcription-recombinase polymerase amplification (RT-RPA) technique combined with a lateral flow strip (LFS). The RT-RPA and RT-RPA-LFS conditions to detect the SMV were optimized using the selected primer set that amplified part of the VPg protein gene. The optimized reaction temperature for the RT-RPA primer and RT-RPA-LFS primer used in this study was 38℃ for both, and the minimum reaction time was 10 min and 5 min, respectively. The RT-RPA-LFS was as sensitive as RT-PCR to detect SMV with 10 pg/µl of total RNA. The reliability of the developed RT-RPA-LFS assay was evaluated using leaves collected from soybean fields. The RT-RPA-LFS diagnostic method developed in this study will be useful as a diagnostic method that can quickly and precisely detect SMV in the epidemiological investigation of SMV, in the selection process of SMV-resistant varieties, on local farms with limited resources.

Isolation and Characterization of a cDNA Encoding Two Novel Heat-shock Factor OsHSF6 and OsHSF12 in Oryza Sativa L.

  • Liu, Jin-Ge;Yao, Quan-Hong;Zhang, Zhen;Peng, Ri-He;Xiong, Ai-Sheng;Xu, Fang;Zhu, Hong
    • BMB Reports
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    • v.38 no.5
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    • pp.602-608
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    • 2005
  • As a crucial transcription factor family, heat-shock factors were mainly analyzed and characterized in tomato and Arabidopsis. In this study, we isolated two putative heat shock factors OsHSF6 and OsHSF12 that interact specifically with heat-shock element (HSE) from Oryza sativa L by yeast one-hybrid method. The full-length cDNA of OsHSF6 and OsHSF12 have 1074bp and 920bp open reading frame (ORF), respectively. Analysis of the deduced amino acid sequences revealed that OsHSF6 was a class A heat shock factor (HSF) with all the conserved sequence elements characteristic of heat stress transcription factor, while OsHSF12 was a class B HSF with C-terminal domain (CTD) lacking of AHA motif. Bioinformatic analysis showed that the sequences and structures of two HSFs' DNA binding domain (DBD) had a high similarity with LpHSF24. The results of RT-PCR indicated OsHSF6 gene was expressed immediately after rice plants exposure to heat stress, and the transcription of OsHSF6 gene accumulated primarily in immature seeds, roots and leaves. However, we did not find the transcription of OsHSF12 gene in different organs and growth periods. Our results implied that OsHSF6 might be function as a HSF regulating early expression of stress genes in response to heat shock, and OsHSF12 might be act as a synergistic factor to regulate the expression of down-stream genes.

Development of a Reporter System Monitoring Regulated Intramembrane Proteolysis of the Transmembrane bZIP Transcription Factor ATF6α

  • Kim, Jin-Ik;Kaufman, Randal J.;Back, Sung Hoon;Moon, Ja-Young
    • Molecules and Cells
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    • v.42 no.11
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    • pp.783-793
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    • 2019
  • When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of $ATF6{\alpha}$ by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human $ATF6{\alpha}$ N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-$hATF6{\alpha}$ deletion variant was cleaved to liberate active transcription activator encompassing GV-$hATF6{\alpha}$ fragment which could translocate into the nucleus. The translocated GV-$hATF6{\alpha}$ fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-$hATF6{\alpha}$(333) represents an innovative tool to investigate regulated intramembrane proteolysis of $ATF6{\alpha}$. It can substitute active pATF6(N) binding motif-based reporter cell lines.

A study on improving the performance of the machine-learning based automatic music transcription model by utilizing pitch number information (음고 개수 정보 활용을 통한 기계학습 기반 자동악보전사 모델의 성능 개선 연구)

  • Daeho Lee;Seokjin Lee
    • The Journal of the Acoustical Society of Korea
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    • v.43 no.2
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    • pp.207-213
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    • 2024
  • In this paper, we study how to improve the performance of a machine learning-based automatic music transcription model by adding musical information to the input data. Where, the added musical information is information on the number of pitches that occur in each time frame, and which is obtained by counting the number of notes activated in the answer sheet. The obtained information on the number of pitches was used by concatenating it to the log mel-spectrogram, which is the input of the existing model. In this study, we use the automatic music transcription model included the four types of block predicting four types of musical information, we demonstrate that a simple method of adding pitch number information corresponding to the music information to be predicted by each block to the existing input was helpful in training the model. In order to evaluate the performance improvement proceed with an experiment using MIDI Aligned Piano Sounds (MAPS) data, as a result, when using all pitch number information, performance improvement was confirmed by 9.7 % in frame-based F1 score and 21.8 % in note-based F1 score including offset.

Pan-serotype reverse transcription loop-mediated isothermal amplification (RT-LAMP) for the rapid detection of foot-and-mouth disease virus (구제역바이러스 신속진단을 위한 pan-serotype reverse transcription loop-mediated isothermal amplification (RT-LAMP) 진단법)

  • Lim, Da-Rae;Park, Yu-Ri;Park, Sun-Young;Kim, Hye-Ryung;Park, Min-Ji;Ku, Bok-Kyung;Nah, Jin-Ju;Ryoo, So-Yoon;Wee, Sung-Hwan;Jeon, Hyo-Sung;Kim, Ji-Jeong;Jeon, Bo-Young;Lee, Hyeong-Woo;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.41 no.1
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    • pp.29-39
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    • 2018
  • In this study, we developed a sensitive and specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid visual detection of foot-and-mouth disease virus (FMDV) circulated in Korea. The RT-LAMP was completed in 40 min at $62^{\circ}C$ and the results of the assay were directly detected by naked eye without any detection process. The assay specifically amplified all 7 serotypes of FMDV RNAs but not amplified other viral and cellular nucleic acids. The sensitivity of the RT-LAMP was $10^2$, $10^3$ and $10^3TCID_{50}/mL$ for serotype O, A and Asia 1 FMDV, respectively, which was comparable to conventional reverse transcription polymerase chain reaction (RT-PCR) and relatively lower than that of real time quantitative RT-PCR (qRT-PCR). Clinical evaluation of the RT-LAMP using different serotypes of Korean and foreign FMDV strains showed a 100% (35/35) agreement with the results of the RT-PCR and qRT-PCR. These results indicated that RT-LAMP assay developed in this study could be a valuable diagnostic method for FMDV monitoring and surveillance.

NMF Based Music Transcription Using Feature Vector Database (특징행렬 데이터베이스를 이용한 NMF 기반 음악전사)

  • Shin, Ok Keun;Ryu, Da Hyun
    • Journal of Advanced Marine Engineering and Technology
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    • v.36 no.8
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    • pp.1129-1135
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    • 2012
  • To employ NMF to transcribe music by extracting feature matrix and weight matrix at the same time, it is necessary to know in advance the dimension of the feature matrix, and to determine the pitch of each extracted feature vector. Another drawback of this approach is that it becomes more difficult to accurately extract the feature matrix as the number of pitches included in the target music increases. In this study, we prepare a feature matrix database, and apply the matrix to transcribe real music. Transcription experiments are conducted by applying the feature matrix to the music played on the same piano on which the feature matrix is extracted, as well as on the music played on another piano. These results are also compared to those of another experiment where the feature matrix and weight matrix are extracted simultaneously, without making use of the database. We could observe that the proposed method outperform the method in which the two matrices are extracted at the same time.