UNIT 12.7 Regulation of the HIV Promoter/Enhancer
Published Online: 1 MAY 2003
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Palmieri, C., Trimboli, F., Scala, G., Quinto, I. and Bressler, P. B. 2003. Regulation of the HIV Promoter/Enhancer. Current Protocols in Immunology. 54:12.7:12.7.1–12.7.18.
- Published Online: 1 MAY 2003
- Published Print: APR 2003
This unit describes adaptations of two molecular techniques that can be used to study the regulation of HIV expression. The first two protocols describe the chloramphenicol acetyltransferase (CAT) assay, in which the CAT reporter gene is attached to an HIV-1 promoter and CAT activity is measured as an indication of the promoter's activity. The basic protocol is rapid, simple, and suited to analyzing multiple samples. An alternate protocol describes an assay for CAT function that involves separating the reaction products by thin-layer chromatography (TLC). The second describes an electrophoretic mobility shift assay for detecting proteins present in cell extracts that can bind to the HIV-1 LTR (long terminal repeat). Such studies are central to current HIV research because it is important to know what agents induce and inhibit (or “down-regulate”) HIV transcription.