Analysis of the polyhedrin gene promoter of theAutographa californicanuclear polyhedrosis virus

Abstract

The polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus was analysed with respect to which sequences are required upstream of the mRNA transcription initiation (CAP) site for efficient promoter activity. Insertions (8, 95 and 785 nucleotides) were made in this region at an EcoR V site between the CAAT- and TATA-like boxes. When these mutations were introduced into the virus they did not affect the activity of the polyhedrin promoter as judged by expression of the beta-galactosidase (lacZ) gene inserted in lieu of the polyhedrin coding sequences. Deletions were made in the promoter which progressively removed sequences upstream from the CAP site. Removal of the TATA motif did not affect lacZ gene expression. A sequence 69 nucleotides upstream to the normal position of the polyhedrin ATG translation initiation codon was sufficient for maximum promoter activity but this was reduced by 90% when only 56 nucleotides upstream remained. The normal CAP site was utilized by each deletion mutant. Promoter activity was undetectable when the CAP site was deleted. The results are discussed in relation to other eukaryotic promoters.

Location and nucleotide sequence of the gene encoding the viral enhancing factor of the Trichoplusia ni granulosis virus

“…However, in all three sequences, there is a mismatch at the invariable T of the ATAAG core late promoter motif. Changes at this position have been shown to eliminate transcription initiation (Possee & Howard, 1987;Rankin et al, 1988;Ooi et al, 1989). A comparison of the deduced amino acid sequence of the VEF with both the NBRF and Swiss-Prot protein databases did not reveal any similarity to known proteins.…”

The p6.5 gene region of a nuclear polyhedrosis virus of Orgyia pseudotsugata: DNA sequence and transcriptional analysis of four late genes

“…2 and 3). This sequence has been implicated as necessary for transcription of baculovirus late genes (Rohrmann, 1986;Possee & Howard, 1987;Rankin et al, 1988). The nucleotide at the -3 position, relative to the translation start codon of all the ORFs, contains a purine base (Fig.…”

RNA editing-like phenomenon in paramyxovirus V gene mRNA observed in insect cells infected with a recombinant baculovirus.

“…Methods for the preparation of an infectious baculovirus DNA, AcRP23.lacZ (Possee & Howard, 1987), have been described previously (Summers & Smith, 1988). AcRP23.lacZ and lipofectin (Gibco) were used for co-transfection with the transfer vector.…”