Studies on the Control Region of the p10 Gene of the Autographa californica Nuclear Polyhedrosis Virus

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Abstract
Summary 5′ deletion mutants of the Autographa californica nuclear polyhedrosis virus very late p10 gene promoter have been prepared and subjected to a transient expression assay in infected Spodoptera frugiperda cells. The control plasmid contained the chloramphenicol acetyltransferase (CAT) reporter gene under the control of the p10 promoter, which was included in a 230 bp sequence upstream from the p10 translation initiation codon. The control plasmid also contained a segment of the hr5 enhancer downstream from the CAT gene. Promoter activity was unaffected by 5′ deletion to position -77, which lies about 11 bp upstream from the p10 cap site. However, deletion of 12 more bp completely eliminated p10 promoter activity. Thus, the 5′ border of the p10 promoter lies downstream from position -77, and the region between positions -77 and -65 contains an element that is important to promoter activity. This is the region that is conserved near the cap sites of late baculovirus genes. Our studies also show that transient expression of CAT under the control of the p10 promoter and hr5 enhancer is higher when transfection occurs prior to infection by virus.