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The adenovirus (AV) backbone for the aquaporin-1 (AQP1) sense and antisense AV constructs was a replication-deficient "first-generation" AV with deletions of the E1 and E3 genes [ 26 ] . This "empty" AV contains the cytomegalovirus (CMV) promoter and bovine growth hormone polyadenylation (bHG) site separated by a polylinker that was used to clone AQP1 DNA as described previously [ 27 ] . A recombinant AQP1 virus was constructed using a plasmid containing the coding sequence for AQP1, pCHIPev [ 28 ] . pCHIPev was digested and the AQP1 insert was subcloned into the shuttle vector pSKAC, creating pSKAC/AQP1.
pSKAC contains map units 0.0-1.
DNA fragments containing AQP1 DNA were liberated from pSKAC after restriction and ligated into adenovirus as described previously [ 27 ] . Human embryonic kidney cells (293 cells) were transfected with ligation mixture and individual viruses were isolated from cell lysates by two consecutive rounds of plaque purification using an agar overlay as described previously [ 27 ] . Individual viruses were amplified in 293 cells and purified over cesium step gradient.