High level expression and structural characterisation of herpes simplex virus type I transcriptional activator VP16 (alpha-trans inducing factor).
We have developed a baculovirus expression system for the rapid and efficient production of large quantities (>10 mg/l) of VP16. The recombinant VP16 binds to a complex of host cell transcription factors and TAATGARAT motif. Secondary structure calculations from circular dichroism measurements indicate a content of 32.0 % alpha-helix and 17.5 % beta-sheet. This is the first structural CD analysis of VP16 which will be very useful for high-throughput assay development and mechanistic studies.
Spatz
A M
AM
Institut für Bakteriologie, Mykologie, und Hygiene, Veterinärmedizinische Universität Wien, Veterinaerplatz 1, Vienna, A-1210, Austria. Martin.Spatz@vu-wein.ac.at
Ali
S A
SA
Auer
M
M
Graf
C
C
Eibl
M M
MM
Steinkasserer
A
A
D016367:Baculoviridae / Q000235:genetics; D002942:Circular Dichroism; D015967:Gene Expression Regulation, Viral; D017405:Herpes Simplex Virus Protein Vmw65 / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000737:chemistry*; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D017405:Herpes Simplex Virus Protein Vmw65; D011994:Recombinant Proteins