Differential transcription of baculovirus late and very late promoters: fractionation of nuclear extracts by phosphocellulose chromatography

• DOI: 10.1128/jvi.69.5.2912-2917.1995
• Publication Date: 2020-01-06T21:30:39Z
• Authors: B Xu; S Yoo; L A Guarino
• Keywords: Polyhedrin; Autographa californica; Transcription; Nuclear Polyhedrosis Virus; RNA polymerase II

An in vitro transcription system based on cytidine-free cassette was developed for the late 39k gene and very polyhedrin of Autographa californica nuclear polyhedrosis virus (AcNPV). Optimization conditions revealed that a preincubation step not required promoters, although essential efficient from an early promoter. The constructs were actively transcribed by extracts prepared AcNPV-infected Spodoptera frugiperda cells at 12 or 36 h postinfection but uninfected infected harvested during phase infection. Transcription promoter fivefold higher than with extract postinfection. 36-h fractionated phosphocellulose chromatography. activity eluted two fractions, 0.3 0.5 M KCl. Both these fractions; however, patterns distinctly different. With fraction, incorporation into transcript approximately 10-fold transcript. Alternatively, construct twofold construct. These results indicate this will be useful purification identification factors discriminate between promoters.