Phosphoramidation Reactions for Nucleic Acids Conjugating with Peptides, Proteins, Chromophores and Biotin Derivaties

• Main Authors: Yi-Jang Chiou, 邱奕璋
• Other Authors: Tzu-Pin Wang
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/99779113424401190072

碩士 === 高雄醫學大學 === 醫藥暨應用化學研究所 === 98 === The original aim of this research was to develop a new phosphoramidation reaction suitable for nucleic acid and small organic compound conjugations which could be exploited to identify amidase ribozymes from RNA pools. An amine-containing biotin derivative (11) was synthesized and used to systematically optimize a previously ineffective and inefficient phosphoramidation reaction method. The improved phosphoramidation reaction increased yields of nucleic acid-11 conjugates up to 80% after 3-hr reactions. Applications of the phosphoramidation reactions to conjugate 96 RNA clones with 11 and studies of the enzymatic activities for these RNA-11 conjugates revealed that all RNA conjugates had no detectable amidase activity. We, however, demonstrated any nucleic acids with a terminal phosphate group were suitable reactants in the new phosphoramidation reactions to conjugate with nucleophile molecules such as derivatives of biotin and fluorescein, proteins and peptides. Polymerase chain reaction (PCR) studies of 11- or fluorescein-tagged DNA primers prepared by the phosphoramidation reactions indicated base-pairing characteristics of the nucleic acid conjugates were substained. This study developed a facile phosphoramidation approach to prepare nucleic acid conjugates with good yields which have implications for broad biomedical applications.