Circular Dichroism Analysis of Insect Cell Expressed Herpes Simplex Virus Type I Single-Stranded DNA-Binding Protein ICP8

We have developed a baculovirus expression system for the rapid and efficient production of large quantities (>5 mg/108cells) of ICP8. The recombinant ICP8 is fully functional and binds to single-stranded DNA. Secondary structure calculations from circular dichroism measurements indicate a conten...

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Bibliographic Details
Published in:Protein expression and purification 1999-06, Vol.16 (1), p.40-46
Main Authors: Spatz, Martin, Ali, Stuart A., Auer, Manfred, Graf, Christine, Eibl, Martha M., Steinkasserer, Alexander
Format: Article
Language:English
Subjects:
Animals
Baculoviridae - genetics
baculovirus expression system
Base Sequence
Cell Line
Circular Dichroism
DNA Primers - genetics
DNA, Single-Stranded - metabolism
DNA-Binding Proteins - chemistry
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Gene Expression
herpes simplex virus
ICP8
In Vitro Techniques
Protein Structure, Secondary
recombinant protein
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Simplexvirus - genetics
Spodoptera
Viral Proteins - chemistry
Viral Proteins - genetics
Viral Proteins - metabolism
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Summary:We have developed a baculovirus expression system for the rapid and efficient production of large quantities (>5 mg/108cells) of ICP8. The recombinant ICP8 is fully functional and binds to single-stranded DNA. Secondary structure calculations from circular dichroism measurements indicate a content of 34.5% α-helix and 15.4% β-sheet. This is the first structural report for ICP8 using CD analysis, which will be very useful for high-throughput assay development and mechanistic studies.
ISSN:1046-5928
1096-0279
DOI:10.1006/prep.1999.1046