Methods for analysis of incorporation and annealing methods for analysis of tRNALys in HIV-1

Abstract

In HIV-1, tRNALys3 serves as the primer for reverse transcription of minus strand strong stop cDNA. During viral assembly, the tRNALys isoacceptors, tRNALys1,2 and tRNALys3, are selectively packaged into the virion. The selectively packaging of tRNA Lys3 facilitates the annealing of tRNALys3 to the viral genome and the initiation of reverse transcription. We describe herein a set of experimental approaches for studying the mechanism by which tRNALys is selectively incorporated into HIV-1 and investigate how primer tRNA Lys3 is annealed to viral genome. The methods described will also help in the analysis of cellular RNAs packaged in the virus particles. © 2009 Humana Press.