Methods for Analysis of Incorporation and Annealing Methods for Analysis of tRNALys in HIV-1

In HIV-1, tRNA^Lys3 serves as the primer for reverse transcription of minus strand strong stop cDNA. During viral assembly, the tRNA^Lys isoacceptors, tRNA^Lys1,2 and tRNA^Lys3 , are selectively packaged into the virion. The selectively packaging of tRNA^Lys3 facilitates the annealing of tRNA^Lys3 to the viral genome and the initiation of reverse transcription. We describe herein a set of experimental approaches for studying the mechanism by which tRNA^Lys 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.