Unit

UNIT 12.7 Regulation of the HIV Promoter/Enhancer

  1. Camillo Palmieri1,
  2. Francesca Trimboli1,
  3. Giuseppe Scala1,
  4. Ileana Quinto1,
  5. Peter B. Bressler2

Published Online: 1 MAY 2003

DOI: 10.1002/0471142735.im1207s54

Current Protocols in Immunology

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.

Author Information

  1. 1

    Department of Clinical and Experimental Medicine, University of Catanzaro, Magna Grecia, Cantazaro, Italy

  2. 2

    National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2003
  2. Published Print: APR 2003

Abstract

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 basic protocol 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.