Tg(Cdh5-RHO/ADRA1,-lacZ)B26-1Mik

The ~200 kbp C57BL/6J mouse bacterial artificial chromosome (BAC) RP23-453P1 was obtained; containing the entire cadherin 5 locus (Cdh5) as well as ~124 kbp of 5' flanking sequences (including the complete Gm32728, Gm39236 and Gm8748 predicted gene loci) and ~33 kbp of 3' flanking sequences (including a 5' portion of the Bean1 locus). Using homologous recombination/BAC recombineering, a 6170 bp Optoalpha1AR-IRES-lacZ-pA construct was inserted into the ATG start site of the BAC Cdh5 gene (replacing the initiation codon of Cdh5). No other loci on the BAC were altered. The 11.6 kbp BAC vector (pBACe3.6) contained a chloramphenicol selection cassette. The Optoalpha1AR fusion protein is a G>t<-coupled bovine rhodopsin (RHO) sequence modified to have its intracellular regions (including carboxy-terminal domains) replaced with those from the G>q<-coupled human ADRA1A. Founder line B26-1 was identified with high effector expression in vascular endothelial cells. (J:314125)