Hairpin DNA probe-based fluorescence assay for detecting palindrome cleavage activity of HIV-1 integrase

Anal Biochem. 2014 Sep 1:460:36-8. doi: 10.1016/j.ab.2014.05.010. Epub 2014 May 23.

Abstract

HIV-1 integrase is a key enzyme for retroviral replication. The integrase can perform a palindrome cleavage reaction. In this work, a hairpin DNA probe with a palindromic DNA sequence mimicking the HIV-1 LTR-LTR junction of the 2-LTR circles in its long stem was designed for fluorescence detection of specific restriction-like cleavage activity of HIV-1 integrase. Results showed that the designed probe could be recognized and cleaved by HIV-1 integrase. The palindrome cleavage reaction can be monitored according to the increase in fluorescent signal. The assay can be applied to real-time detection of palindrome cleavage of HIV-1 integrase with advantages of simplicity, high sensitivity, and specificity.

Keywords: Fluorescent probe; HIV-1; Integrase; Palindrome cleavage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Probes / genetics*
  • DNA Probes / metabolism*
  • Enzyme Assays / methods*
  • HIV Integrase / metabolism*
  • HIV Long Terminal Repeat
  • HIV-1 / enzymology*
  • HIV-1 / genetics
  • Inverted Repeat Sequences*
  • Spectrometry, Fluorescence

Substances

  • DNA Probes
  • HIV Integrase
  • p31 integrase protein, Human immunodeficiency virus 1