Analysis of cysteine protease inhibitor gene (BmCPI) promoter activity in silkworms using bac-to-bac baculovirus systems

Abstract

A cysteine protease inhibitor of Bombyx mori (BmCPI) plays an important role in pupation, molting, and dissociation of tissues. The present study identified and analyzed the BmCPI promoter region to better understand its functional regulatory mechanisms. Eight promoter fragments of different lengths were analyzed using an improved Bac-to-Bac expression system. Luciferase activities were investigated both in BmE cells and larval organisms after infection with the Bac-to-Bac system, and similar changes in activity were observed in both models. Strong activity was detected in the longest promoter (2005 bp, -1969 to +36), and activity changed significantly with truncation of promoter length. An electrophoretic mobility shift assay showed that the promoter region from -32 to +6 bp played a critical role in activating the downstream gene promoter element, where some potential elements were also predicted by informatics tools. The findings offer a basic reference for the mechanism of transcriptional regulation of BmCPI.

Sex‐specific expression and DNA methylation in a species with extreme sexual dimorphism and paternal genome elimination

“…Putative promoter regions were defined as 2000 bp upstream from each gene. We chose this size based on previous research (Sinha et al, 2006; Tian et al, 2018). We excluded promoters which overlap any other genomic feature using bedtools (Quinlan & Hall, 2010).…”