• Bulletin of the Korean Chemical Society
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• 제29권5호
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• pp.943-947
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• 2008

A phosphate-specific and fluorescent probe was prepared for label-free phosphatase assays based on fluorescence polarization. By using the probe, dephosphorylation reactions of DNA and protein substrates by calf intestinal alkaline phosphatase (CIP) could effectively be monitored in real-time. Since this assay method does not require additional materials such as labeled substrates and phosphospecific antibodies to obtain fluorescence polarization signals, it is simple, cost-effective, and expected to be useful not only for measuring activity of phosphatases but also for high-throughput screening of phosphatase inhibitors.

• 생명과학회지
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• 제29권9호
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• pp.949-954
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• 2019

세포막 단백질의 topology는 막단백질의 중요한 특징이다. 우리는 이전에 C4orf32단백질을 클로닝 하였으나, 이 단백질의 세포내 위치나 topology는 알지 못했다. 이번 연구를 통해 C4orf32는 세포내에서 소포체에 위치되는 막단백질임을 알게 되었다. C4orf32의 topology를 알기 위해 protease protection assay, fluorescence protease protection (FPP) assay, FRB/rapamycin/FKBP system을 활용하였다. Protease protection assay와 FPP assay를 적용한 결과 C-말단에 GFP를 붙인 C4orf32-GFP의 경우 GFP가 소포체의 세포질 표면에 위치함을 확인할 수 있었다. 또한, FRB/rapamycin/FKBP시스템을 이용한 실험에서 rapamycin이 처리되지 않은 경우는 mRFP-FKBP가 세포질에 위치하다가 rapamycin이 처리되면 C4orf32-GFP-FR가 위치하는 소포체로 이동함을 확인할 수 있었다. 이러한 사실은 C4orf32의 C-말단이 소포체의 세포질쪽 면에 위치한다는 사실을 말해준다. 이러한 연구를 통해 C4orf32는 Type I 소포체 막단백질에 속한다는 사실을 확인할 수 있었다.

• The Plant Pathology Journal
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• 제38권6호
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• pp.665-672
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• 2022

Cymbidium mosaic virus (CymMV) is one of economically important viruses that cause significant losses of orchids in the world. In the present study, a reverse transcription recombinase polymerase amplification (RT-RPA) assay combined with a lateral flow immunostrip (LFI) assay was developed for the detection of CymMV in orchid plants. A pair of primers containing fluorescent probes at each terminus that amplifies highly specifically a part of the coat protein gene of CymMV was determined for RT-RPA assay. The RT-RPA assay involved incubation at an isothermal temperature (39℃) and could be performed rapidly within 30 min. In addition, no cross-reactivity was observed to occur with odontoglossum ringspot virus and cymbidium chlorotic mosaic virus. The RT-RPA with LFI assay (RT-RPA-LFI) for CymMV showed 100 times more sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR). Furthermore, the RT-PCR-LFI assay demonstrated the simplicity and the rapidity of CymMV detection since the assay did not require any equipment, by comparing results with those of conventional RT-PCR. On-site application of the RT-RPA-LFI assay was validated for the detection of CymMV in field-collected orchids, indicating a simple, rapid, sensitive, and reliable method for detecting CymMV in orchids.

• Bulletin of the Korean Chemical Society
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• 제34권1호
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• pp.31-32
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• 2013

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.315-319
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• 2003

A novel method is developed to yield virus titers in 10 h, is easy to .perform using 96-well plates, and applicable to both any Autographa californica nucleopolyhyderovirus (AcNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV)-based recombinant baculovirus. This assay uses an antibody to a DNA-binding protein to detect the infected cells via immune-staining. The titer is determined by counting foci produced due to infection of virus under a fluorescent microscopy. The required incubation period was shortened considerably because infected cells expressed viral antigens at the post infection time of 4 h. Therefore, 10 hours were enough to estimate the virus titer including virus infection time, insect cell culture, and estimation of virus titer.

• Macromolecular Research
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• 제17권4호
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• pp.232-239
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• 2009

This paper describes the simple nanopatterning of proteins on polyelectrolyte surfaces using microcontact printing with a nanopatternable, hydrophilic composite nanomold. The composite nanomold was easily fabricated by blending two UV-curable materials composed of Norland Optical Adhesives(NOA) 63 and poly(ethylene glycol) dimethacrylate(PEG-DMA). NOA 63 provided stable nanostructure formation and PEG-DMA induced high wettability of proteins in the nanomold. Using the composite mold and functionalized surface with polyelectrolytes, the fluorescent, isothiocyanate-tagged, bovine serum albumin(FITC-BSA) was successfully patterned with 8 nm height and 500 nm width. To confirm the feasibility of the protein assay on a nanoscale, a glycoprotein-lectin assay was successfully demonstrated as a model system. As expected, the lectins correctly recognized the nano-patterned glycoproteins such as chicken ovalbumin. The simple preparation of composite nanomold and functionalized surface with a universal platform can be applied to various biomolecules such as DNA, proteins, carbohydrates, and other biomolecules on a nanoscale.

• 대한의생명과학회지
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• 제8권4호
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• pp.269-273
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• 2002

Dengue fever (DF) is an acute febrile illness caused by dengue viruses in the family Flaviviridae, genus Flavivirus. DF has so far posed any problem in Korea, however it has been recently believed to be associated with oversea's traveler infected with dengue virus. Antibody titers of sera from DF patients against dengue virus were measured by indirect immunofluorescence assay (IFA) and plaque reduction neutralization test (PRNT), including the haematologic test. Three of patients with DF showed highly fluorescent and neutralizing antibody titers by IFA and PRNT assay. Two of them showed higher, remarkably. Meanwhile, one of them was tested and resulted in severe tirombocytopenia, elevated serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) activities as well as mild leucopenia, increased monocytes and basophils and depressed lymphocytes in haematological differential count.

• 센서학회지
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• 제14권5호
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• pp.308-312
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• 2005

We present a microfluidic chip designed for the detection of antibody by using quantum dots fluorescence and a microbead-based assay. A custom designed PDMS microfluidic chip with multi-layer channel is utilized for capturing microbeads; antibody injection into each micro-well; QD injection; and fluorescence detection. The experiment using the fabricated microfluidic chip has been performed on solutions with various concentrations of antibody and has shown correlated fluorescent intensities.

• 생명과학회지
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• 제18권9호
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• pp.1312-1315
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• 2008

DAPI 염색을 이용하여 세포사멸 유도물질의 초고속스크리닝에 적합한 protocol을 제작하였다. Liquid handler에서 dispensing과 multilabel counter에서의 상대적인 fluorescent intensity를 측정하는 과정을 자동화한 protocol을 제작하였고, 이의 과정은 매우 효율적으로 가동되었음을 확인하였다. DAPI에 binding된 DNA의 intensity를 측정하는 조건검토에서 가장자리의 36개를 reading하지 않는 것이 edge effect를 줄일수 있는 것을 확인하였고, 0.1 sec reading으로 시간을 절약할 수 있었다. 이의 결과를 효과적으로 이용할 경우, 세포사멸 유도물질의 초고속스크리닝 또는 이에 적합한 스크리닝이 가능함을 확인하였다.

• 한국축산식품학회지
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• 제35권3호
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• pp.382-388
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• 2015

A duplex real-time polymerase chain reaction (PCR) based assay for the detection of porcine and horse meat in sausages was designed by using EvaGreen fluorescent dye. Primers were selected from mitochondrial 12S rRNA and 16S rRNA genes which are powerful regions for identification of horse and porcine meat. DNA from reference samples and industrial products was successfully extracted using the GIDAGEN® Multi-Fast DNA Isolation Kit. Genomes were identified based on their specific melting peaks (Mp) which are 82.5℃ and 78℃ for horse and porcine, respectively. The assay used in this study allowed the detection of as little as 0.0001% level of horse meat and 0.001% level of porcine meat in the experimental admixtures. These findings indicate that EvaGreen based duplex realtime PCR is a potentially sensitive, reliable, rapid and accurate assay for the detection of meat species adulterated with porcine and horse meats.