HIV Reverse Transcriptase
Human Immunoeficiency Virus (HIV) Reverse Transcriptase is an RNA directed DNA polymerase which can synthesize a complementary DNA strand initiating from a primer using either RNA or single-stranded DNA as a template.
Description:
· Provides an excellent target for evaluating antiviral agents or inhibitors (1,2).
· Catalyzes error-prone synthesis on DNA and RNA templates - ideal for introducing random mutations (3).
Unit Definition: One unit is the amount of enzyme required to incorporate 1 nmol of labeled dTTP into acid-insoluble material in 10 min at 37°C.
Storage Conditions: Store at –20°C.
Storage Buffer: 20 mM potassium phosphate (pH 7.1), 1 mM dithiothreitol, 0.02% (v/v) Triton™X-100 and 50% (v/v) glycerol.
Assay Conditions: 50 mM Tris-HCl (pH 8.6 at 22°C), 10 mM MgCl2, 40 mM KCl, 0.5 mM [³H]dTTP and 0.4 mM poly(A)·(dT)12-18. Incubation is at 37°C for 10 min in a reaction volume of 50 µl.
Quality Control: All preparations are assayed for contaminating endonuclease and exonuclease activities.
References:
1. Hirsch, M. S., Kaplan, J. C. (1985) Ann. Intern. Med. 103, 750-755. Review.
2. Tisdale, M. et al (1989) J. Antimicrob Chemother. 23, 47-54.
3. Williams, K. J., Loeb, L. A. (1992) Curr. Top. Microbiol. Immunol. 176, 80-165.