Dual-Reporter Phenotypic Assay for Human Immunodeficiency Viruses

Pairwise Growth Competition Assay for Determining the Replication Fitness of Human Immunodeficiency Viruses

In vitro fitness assays are essential tools for determining viral replication fitness for viruses such as HIV-1. Various measurements have been used to extrapolate viral replication fitness, ranging from the number of viral particles per infectious unit, growth rate in cell culture, and relative fitness derived from multiple-cycle growth competition assays. Growth competition assays provide a p...

Dual-reporter assay using two secreted luciferase genes.

Luciferase is used as a reporter enzyme to estimate gene expression in prokaryotic or eukaryotic cells because the amount of luciferase present is correlated with light intensity in the presence of excess luciferin (1). Furthermore, its sensitivity and range of linear responses are superior to those of other typical reporters (2). When the luciferase gene containing a target promoter region is ...

A Dual Reporter Splicing Assay Using HaloTag-containing Proteins

To evaluate the effects of genetic variations on mRNA splicing, we developed a minigene-based splicing assay using reporter genes encoding luciferase and the multifunctional HaloTag protein. In addition to conventional RT-PCR analysis, splicing events can be monitored in this system using two parameters: luciferase activity and signals derived from HaloTag-containing proteins bound to a fluores...

MOLECULAR BIOLOGY OF HUMAN IMMUNODEFICIENCY VIRUSES: HIV-l AND HIV-2

Tat-independent replication of human immunodeficiency viruses.

The replication of human immunodeficiency retroviruses involves a complex series of events that is regulated at both transcriptional and posttranscriptional levels. The tat gene product is a potent trans-activator of viral transcription and therefore an attractive target for the development of antiviral drugs. Tat-defective HIV-1 proviral DNA clones have been shown previously to be replication ...