Iris Lee with Prof. Rebecca Montgomery, Pei-Yin Lim & Shaun Falk

UW-Madison Department of Biochemistry, Institute for Molecular Virology

Seventy to ninety percent of adults in the United States are infected with human herpes simplex virus type 1 (HSV-1).  HSV-1 causes a wide range of syndromes, varying from cold sores to life-threatening infections.  In order to develop effective treatment against HSV-1, we need to understand its infection process, which can be accomplished by studying the phenotypes of virus with mutated envelope protein.  Glycoprotein B (gB) is an essential viral envelope protein because it mediates virus fusion to the host cell.  However, it is difficult to engineer HSV-1 genes directly due to the large size of its genome (150 kbps).  Therefore the first step in making virus with mutated gB protein is to clone the gB gene (2.8 kbps) into an expression plasmid.  To this end, we successfully amplified gB gene from a small amount of HSV-1 DNA by PCR.  However, 2 attempts to clone this PCR gB gene fragment into an expression vector failed.  This failure may be due to mutation at the end of the fragment commonly occurring in PCR.  Nevertheless, we obtained a wildtype gB gene expression vector from a friendly lab and we successfully inserted a marker (his-tag) in front of this gB gene for the purpose of monitoring its expression and function.