Evolution of a Deoxyribozyme Ligase

描述（由申请人提供）：DNA作为稳定催化剂的潜力可能使该分子比核酶更成功地作为分子生物学、生物技术、纳米技术和临床诊断的工具。此外，比较催化RNA结构，这是自然发生的，更频繁，和DNA结构，导致催化基序将被证明是至关重要的，在揭示生物催化剂的基本特征。为了进一步深入了解核酸酶的进化和结构与功能，本提案的目标是：1）使用逐步进化程序从随机序列进化脱氧核酶连接酶，2）优化脱氧核酶催化的连接酶链式反应（LCR），一种通过连接底物和模板的变性和退火循环来指数扩增靶DNA的方法，和3）使用脱氧核酶催化的LCR检测基因突变，例如BRCA 1、BRCA 1的点突变和引起β-地中海贫血的点突变。

DESCRIPTION (provided by applicant): The potential of DNA as a stable catalyst may give this molecule more success than ribozymes as tools for molecular biology, biotechnology, nanotechnology, and clinical diagnostics. Furthermore, comparison of catalytic RNA structures, which occurs naturally and more frequently, and DNA structures that result in catalytic motifs will prove crucial in revealing fundamental features of biocatalysts. To further gain insight into evolution and the structure and function of nucleic acid enzymes, this proposal will aim to 1) evolve a deoxyribozyme ligase from random sequences using stepwise evolution procedures, 2) optimize a deoxyribozyme-catalyzed ligase chain reaction (LCR), a method for the exponential amplification of target DNA through cycles of denaturing and annealing of ligation substrates and template, and 3) detect genetic mutations using the deoxyribozyme-catalyzed LCR, such as point mutations of BRCA1, BRCA1, and ones that give rise to beta-thalassemia.