Photoactivated immobilization of single-stranded DNAs on a psoralen-functionalized surface under low pH conditions

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• 作者：Junhyoung Ahn (12)
  Ju Young Byun (34)
  Yong-Beom Shin (12)
  Min-Gon Kim (45) mkim@gist.ac.kr
• 关键词：DNA immobilization &#8211 ; Psoralen &#8211 ; pH effect &#8211 ; DNA photopatterning &#8211 ; DNA ; protein photocrosslinking
• 刊名：BioChip Journal
• 出版年：2012
• 出版时间：June 2012
• 年：2012
• 卷：6
• 期：2
• 页码：174-183
• 全文大小：390.9 KB
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• 作者单位：1. Nanobiotechnology Major, School of Engineering, University of Science & Technology (UST), Daejeon, 305-806 Korea2. Immune Therapy Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), Daejeon, 305-806 Korea3. Department of Fine Chemical Engineering & Applied Chemistry, Chungnam National University, Daejeon, 305-764 Korea4. Advanced Photonics Reseach Institue, Gwangju Institute of Science & Technology, Gwangju, 500-712 Korea5. Department of Photonics & Applied Physics, Gwangju, 500-712 Korea
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  
• 出版者：The Korean BioChip Society, in co-publication with Springer Verlag GmbH
• ISSN：2092-7843

文摘

Single stranded DNA was photo-patterned on a psoralen-functionalized surface and conjugated with a psoralen-modified enzyme. Photoreactions between the single-stranded DNA and psoralen-functionalized materials were found to be significantaly influenced by pH, showing high efficientcy only under acidic conditions. Specifically, the photopattering of single-stranded DNA probes, such as polythymine, polyadenine, polycytosine, and polyguanine, on a psoralen-functionalized amino-dextran surface under various pH conditions (pH 3.36–8.80) showed that the polythymine probes had the highest immobilization efficiency at pH 3.36. The effect of pH was further confirmed by photopatterning the single-stranded DNA probes using the photomask. While almost no pattern was observed on the psoralen surface at pHs greater than 5.5, much more visible patterns were observed as the pH was decreased below 5.5. As a result of the irradiation of polyT₁₅ and polyA₁₅-labeled DNA probes, the photopatterns formed by the irradiation of polythymine probes were found to be associated with the cross-linking between the thymine and the psoralen moieties. Finally, additional studies showed that psoralen could be used as the reactive center in the photo-induced cross-linking reaction between DNA and the model protein, horseradish peroxidase (HRP).