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

The Varkud Satellite (VS) ribozyme is the largest of the “small” nucleolytic
ribozymes and is the only one for which there are no high resolution crystal structures
available. The VS ribozyme comprises a catalytic domain and a substrate domain. The
catalytic 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. This
naturally brings the cleavage site in close proximity to the A730 loop in helix VI. The
adenines within the A730 loop are very crucial to the cleavage reaction and any
substitution causes a major decrease in the cleavage activity of the ribozyme.
This study is aimed at designing and producing a variant of the Varkud Satellite
ribozyme that consists of multiple parts that can be used for detailed studies of
ribozyme kinetics and assembly.

Identiferoai:union.ndltd.org:WATERLOO/oai:uwspace.uwaterloo.ca:10012/4710
Date January 2009
CreatorsChu, Allen Wing Ho
Source SetsUniversity of Waterloo Electronic Theses Repository
LanguageEnglish
Detected LanguageEnglish
TypeThesis or Dissertation

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