• Title/Summary/Keyword: direct transcription method

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Direct Multiplex Reverse Transcription-Nested PCR Detection of Influenza Viruses Without RNA Purification

  • Song, Man-Ki;Chang, Jun;Hong, Yeong-Jin;Hong, Sung-Hoi;Kim, Suhng-Wook
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1470-1474
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    • 2009
  • This paper describes the development a of direct multiplex reverse transcription-nested polymerase chain reaction (PCR) method, devised for simultaneous detection and typing of influenza viruses. This method combines the direct reverse transcription reaction without RNA purification with the enhancement of sensitivity and specificity of nested PCR. The method successfully detected three major human influenza viruses: influenza virus A subtype 1 (H1N1) and subtype 3 (H3N2), and influenza B virus (B). The minimum number of virus particles (pfu/ml) necessary for detection in spiked saliva samples was 200 (H1N1), 140 (H3N2), and 4.5 (B). The method's sensitivity and simplicity will be convenient for use in clinical laboratories for the detection and subtyping of influenza and possibly other RNA viruses.

One-Step Selection of Artificial Transcription Factors Using an In Vivo Screening System

  • Bae, Kwang-Hee;Kim, Jin-Soo
    • Molecules and Cells
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    • v.21 no.3
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    • pp.376-380
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    • 2006
  • Gene expression is regulated in large part at the level of transcription under the control of sequence-specific transcriptional regulatory proteins. Therefore, the ability to affect gene expression at will using sequencespecific artificial transcription factors would provide researchers with a powerful tool for biotechnology research and drug discovery. Previously, we isolated 56 novel sequence-specific DNA-binding domains from the human genome by in vivo selection. We hypothesized that these domains might be more useful for regulating gene expression in higher eukaryotic cells than those selected in vitro using phage display. However, an unpredictable factor, termed the "context effect", is associated with the construction of novel zinc finger transcription factors--- DNA-binding proteins that bind specifically to 9-base pair target sequences. In this study, we directly selected active artificial zinc finger proteins from a zinc finger protein library. Direct in vivo selection of constituents of a zinc finger protein library may be an efficient method for isolating multi-finger DNA binding proteins while avoiding the context effect.

Determination of Initial Conditions for Tetrahedral Satellite Formation

  • Yoo, Sung-Moon;Park, Sang-Young
    • Journal of Astronomy and Space Sciences
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    • v.28 no.4
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    • pp.285-290
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    • 2011
  • This paper presents an algorithm that can provide initial conditions for formation flying at the beginning of a region of interest to maximize scientific mission goals in the case of a tetrahedral satellite formation. The performance measure is to maximize the quality factor that affects scientific measurement performance. Several path constraints and periodicity conditions at the beginning of the region of interest are identified. The optimization problem is solved numerically using a direct transcription method. Our numerical results indicate that there exist an optimal configuration and states of a tetrahedral satellite formation. Furthermore, the initial states and algorithm presented here may be used for reconfiguration maneuvers and fuel balancing problems.

Direct Analysis of the Transcription of Escherichia coli rnpB Gene Harbored in a Multicopy Plasmid during Bacterial Growth

  • Park, Jeong-Won;Jung, Young-Hwan;Park, Bo-Hyun;Jeoung, Yeon-Hee;Lee, Young-Hoon
    • BMB Reports
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    • v.29 no.3
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    • pp.221-224
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    • 1996
  • To examine the growth-phase dependent control of Escherichia coli rnpB gene we used a combination of Northern analysis for RNA determination and Southern analysis for plasmid DNA determination. The relative amounts of metabolically unstable transcript derived from the internally deleted rnpB gene harbored on a multicopy plasmid as well as the relative plasmid contents were measured by Northern analysis and Southern analysis, respectively, of total nucleic acids from E coli cells containing the plasmid. The relative transcription activity of the rnB was represented by a ratio of the relative amount of the transcript to that of the plasmid DNA during bacterial growth. The rnpB transcription increased rapidly with time during exponential growth, but started to decrease before the transition period of an exponential growing cell culture into the stationary phase. Although the expression pattern was similar to the changes of ${\beta}-galactosidase$ activity expressed from the lysogenic strain carrying the chromosomal rnpB-lacZ fusion which were shown in a previous work, the present data appears to represent a more actual growth-phase control of the rnpB transcription than the previous data by the ${\beta}-galactosidase$ assay. In addition the present method described for a direct analysis of both RNA and plasmid DNA provides a rapid and efficient method that can applied to an examination of transcription control by using a multicopy plasmid.

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Quantitation of Hepatitis C Viral RNA Using Direct CRT-PCR

  • Park, Young-Suk;Lee, Kyung-Ok;Oh, Moon-Ju;Chai, Young-Gyu
    • BMB Reports
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    • v.30 no.3
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    • pp.234-236
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    • 1997
  • Chronic hepatitis C virus (HCV) infection is associated with the rapid development of cirrhosis and hepatocellular carcinoma. It has been reported that the amount of HCV RNA may be correlated with the progression of hepatitis and may be a prognostic marker for treatment of HCV patients. The direct detection of HCV RNA by reverse transcription-polymerase chain reaction (RT-PCR) is widely used to determine the presence of circulating virions. The most relevant limit of this approach is the lack of quantitative information about the viral titer. In the present study, we developed the method for HCV quantitation using competitive reverse transcription (CRT)-PCR using the deleted HCV standard. The serially diluted standard was added in titrated amounts to the target HCV RNA. The mixture was then reverse transcribed and amplified in the same reaction tube. The methods were evaluated using over 110 HCV-PCR positive samples in Koreans. About 59% of the samples were judged to contain $10^{5}-10^{6}$ copies of HCV RNA in 1 ml of serum.

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A Study on Optimal Earth-Moon Transfer Orbit Design Using Mixed Impulsive and Continuous Thrust (순간 및 연속 추력을 이용한 지구-달 최적 전이궤도 설계에 관한 연구)

  • No, Tae-Soo;Jeon, Gyeong-Eon
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.38 no.7
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    • pp.684-692
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    • 2010
  • Based on the planar restricted three body problem formulation, optimized trajectories for the Earth-Moon transfer are obtained. Mixed impulsive and continuous thrust are assumed to be used, respectively, during the Earth departure and Earth-Moon transfer/Moon capture phases. The continuous, dynamic trajectory optimization problem is reformulated in the form of discrete optimization problem by using the method of direct transcription and collocation, and then is solved using the nonlinear programming software. Representative results show that the shape of optimized trajectory near the Earth departure and the Moon capture phases is dependent upon the relative weight between the impulsive and the continuous thrust.

Direct Growth of Patterned-Graphene Using PVP Nanowire Shadow Mask (PVP 나노와이어를 활용한 패턴된 그래핀의 직성장)

  • Eunho Lee;Daesuk Bang
    • Journal of Adhesion and Interface
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    • v.24 no.4
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    • pp.120-123
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    • 2023
  • Graphene, with its exceptional mechanical and electrical properties, has gained significant attention from researchers due to its superior characteristics compared to conventional materials. However, the application of graphene in electronic devices requires a crucial transcription and patterning process, which often introduces numerous defects, substantially impairing its properties. To overcome this limitation and unlock the full potential of graphene for commercial use, there have been various efforts to develop integrated processes for transcription and patterning. In this study, we present a novel growth method that simultaneously achieves precise patterning using polymer nanowires as masks, allowing for the direct growth of graphene. This innovative approach holds promise for realizing advanced electronic components based on nanomaterials in the future.

An study of transcription by processing conditions of Direct Surface Forming Method (DSF의 성형조건 변화가 전사성에 미치는 영향에 관한 연구)

  • 조광환;윤경환
    • Proceedings of the Korean Society for Technology of Plasticity Conference
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    • 2003.10a
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    • pp.221-224
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    • 2003
  • Recently, the market share of the thin-film-transistor liquid-crystalline-display (TFT-LCD) is growing rapidly in display device market. The backlight unit is used as a light source of TFT-LCD module. A light-guide is one of several important components of backlight unit. The manufacturing technology and optical system design of the light guide is very sensitive to quality and cost of the TFT-LCD module. In the present study a new manufacturing method which is called as direct surface forming(DSF) has been tested under various conditions. The result of this test, V-groove pattern shows different shapes depends on the temperature of mold surface, contact time of mold and depth of V-groove.

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Enhanced In Vitro Protein Synthesis Through Optimal Design of PCR Primers

  • Ahn Jin-Ho;Son Jeong-Mi;Hwang Mi-Yeon;Kim Tae-Wan;Park Chang-Kil;Choi Cha-Yong;Kim Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.355-359
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    • 2006
  • The functional stability of mRNA is one of the crucial factors affecting the efficiency of in vitro translation. As the rapid degradation of mRNA in the cell extract (S30 extract) causes early termination of the translational reactions, extending the mRNA half-life will improve the productivity of the in vitro protein synthesis. Thus, a simple PCR-based method is introduced to increase the stability of mRNA in an S30 extract. The target genes are PCR-amplified with primers designed to make the ends of the transcribed mRNA molecule anneal to each other. When compared with normal mRNA, the mRNA with the annealing sequences resulted in an approximately 2-fold increase of protein synthesis in an in vitro translation reaction. In addition, sequential transcription and translation reactions in a single tube enabled direct protein expression from the PCR-amplified genes without any separate purification of the mRNA.

Detection of Soybean mosaic virus by Reverse Transcription Loop-mediated Isothermal Amplification (Reverse transcription Loop-mediated isothermal amplification을 이용한 Soybean mosaic virus의 진단)

  • Lee, Yeong-Hoon;Bae, Dae-Hyeon;Kim, Bong-Sub;Yoon, Young-Nam;Bae, Soon-Do;Kim, Hyun-Joo;Mainali, Bishwo P.;Park, In-Hee;Lee, Su-Heon;Kang, Hang-Won
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.315-320
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    • 2015
  • Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I under daylight and UV light. Optimal reaction condition was at $58^{\circ}C$ for 60 min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.