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Key words: AcNP V/plO promoter/CA T expression Analysis of the Promoter of the Autographa californica Nuclear Polyhedrosis Virus pl0 Gene

By Weyer and Robert D. Possee


Functional analyses of the pl0 gene promoter from the Autographa californica nuclear polyhedrosis virus (AcNPV) were performed by progressively deleting the 230 nucleotides upstream from the pl0 coding sequences towards the ATG codon. Truncated promoter sequences retaining the full 5 ' non-coding leader of pl0 were inserted in front of the chloramphenicol acetyltransferase (CAT) gene, and promoter activity in transfected AcNPV-infected cells was measured using the transient CAT expression assay. The removal of sequences to a position 101 nucleotides upstream from the pl0 ATG did not affect the level of CAT expression. Deletion of a further 13 nucleotides reduced CAT expression by three- to fourfold, but the removal of three more nucleotides, which deleted most of the baculovirus very late gene transcription consensus sequence, almost completely abolished activity. The removal of the TATA motif had no effect on the level of transient expression. We conclude that a sequence of about 101 nucleotides upstream from the ATG codon of p l0 is sufficient for high level promoter activity in this transient system. Throughout the very late phase of infection of insect cells with the Autographa californic

Year: 1988
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