Demonstration of a Ribozyme in Epsilon Domain of Hepatitis B Virus RNA
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The epsilon domain of Hepatitis B virus plays a crucial role in encapsidation of viral pregenomic RNA and its partial NMR structure has been determined. However, we recently described a potassium-dependent ribonucleolytic activity associated with this region, so that a 53 nt long RNA containing the epsilon domain could release itself and cleaved other RNAs. We describe here the experimental methodologies for setting up the reactions and outline a general strategy for initial demonstration of this self-cleaving ribozyme activity.
Key wordsHBV Epsilon Ribozyme Potassium
S.G. acknowledges partial support of this work by Department of Science and Technology (Government of India) grant no. SR/FT/L-100/2004. Department of Biotechnology-BUILDER facility in University of Calcutta was used for Phosphorimager analysis. D.C. was supported by research fellowships from CSIR (9/28(664)/2006 EMR-I) and DBT (BT/01/CEIB/09/VI/10), Govt. of India.
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