Abstract
Objective: The development of a human immunodeficiency virus (HIV)–1 promoter reporter system is critical for investigating transcription activity of the virus. We designed a modified overlap extension PCR method for single round site-directed mutagenesis of the long terminal repeat (LTR) segment of HIV-1. Methods: The procedure consists of overlap extension PCR, DpnI digestion, and product transformation. Wild-type and serial sequence deletion clones of HIV-1 LTR were constructed. Basal transcription activity was also measured. Results: The overall efficiency for obtaining the desired products was 53.2%. The reporter assay indicated the importance of the second NF-ĸB cis-element and surrounding regions in mediating the regulation of HIV-1 transcription. Conclusion: The technique we investigated is suitable for highthroughput functional studies of virus-host interaction.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
