Analysis of calsequestrin gene expression using green fluorescent protein in Caenorhabditis elegans

Abstract

The calsequestrin gene of Caenorhabditis elegans is expressed in body-wall muscle cells during muscle development. In order to study the body-wall muscle specific regulation of the calsequestrin gene expression, approximately 2 kb upstream sequences of the calsequestrin gene were analyzed, Transcriptional fusion constructs utilizing green fluorescent protein as a reporter gene were made and microinjected to produce germ-line transformed transgenic C, elegans, The expression of green fluorescent protein was observed in the body-wall muscles of live transgenic animals under fluorescence microscopy, Deletion analyses of upstream sequences have revealed a putative promoter sequence and a regulatory element which appeared to enhance reporter gene expression. Both sequence elements are juxtaposed to constitute a 260 bp regulatory region approximately 260 bp upstream from the putative translational initiation codon, Several possible binding sites for transcription factors were identified including the sites for Wi and NF-W2, a muscle specific zinc finger transcription factor, and an ubiquitous enhancer binding protein, respectively, Interestingly, this region also contains a 20 bp sequence element identical to those found in the mouse dystrophin gene, which suggests a possible role of this regulatory region in muscle specific gene regulation.