Mouse connexin40: gene structure and promoter analysis.

摘要: A family of related connexin genes encodes the subunit gap junction proteins that form intercellular channels in different tissues. Connexin40 (Cx40) is one these proteins, and it exhibits limited expression only a few cells cardiovascular system. To begin to analyze Cx40 expression, we isolated 3.3-kb rat cDNA by hybridization screening bacteriophage library prepared from BWEM corresponding mouse genomic clones bacterial artificial chromosome library. Restriction mapping, sequencing, comparison showed gene contained short first exon, an 11.4-kb intron, second exon containing complete coding region 3'-UTR. Exon I 1 base differed between mouse. Primer extension experiments yielded single band confirmed position transcriptional start site. We obtained 1.2 kb sequence 5' site 400 bp 3' I. was closely preceded consensus TATA box. The flanking sequences number potential transcription factor binding sites (including AP-1, AP-2, SP1, TRE, p53). identify regulatory elements promoter region, series DNA deletion fragments prepared, subcloned adjacent luciferase reporter gene, used for transient transfections BWEM, SHM, N2A cells. resulting activity determinations suggested area 300 acted as basal there strong negative element +100 +297.