Preparation of a Multi-Part Varkud Satellite Ribozyme Variant for Kinetics Studies

Ribozyme;RNA;Chemistry

Chu, Allen Wing Ho

University of Waterloo

关键词: Ribozyme;    RNA;    Chemistry;   

Others  :  https://uwspace.uwaterloo.ca/bitstream/10012/4710/1/chu_allen.pdf

瑞士|英语

来源: UWSPACE Waterloo Institutional Repository

【 摘 要 】

The Varkud Satellite (VS) ribozyme is the largest of the ;;small” nucleolyticribozymes and is the only one for which there are no high resolution crystal structuresavailable. The VS ribozyme comprises a catalytic domain and a substrate domain. Thecatalytic domain includes five helices that interact with the stem-loop substrate.The substrate is docked within a cleft that is formed by helices II and VI. Thisnaturally brings the cleavage site in close proximity to the A730 loop in helix VI. Theadenines within the A730 loop are very crucial to the cleavage reaction and anysubstitution causes a major decrease in the cleavage activity of the ribozyme.This study is aimed at designing and producing a variant of the Varkud Satelliteribozyme that consists of multiple parts that can be used for detailed studies ofribozyme kinetics and assembly.