Luciferase assay for the functional analysis of HIV tat and LTR in gene expression

A series of LTR deletion-directed Luc expression plasmids were constructed and transfected into Jurkat cells with or without a Tat-producing expression vector. Luciferase activity was measured to analyze the effect of Tat and LTR on gene expression. The results show that there are negative regulatory elements in the LTR upstream from the -158 position. It is demonstrated that trans-activation of. Tat may enhance significantly the level of LTR-directed gene expression. The data suggest that the enhancer sequences in LTR and the binding protein NF- kappa B may play an important role in both high-level LTR mediated transcription and Tat trans-activation.