Isolation, characterization, and expression in Escherichia coli of a cDNA encoding rat heme oxygenase-2.

摘要: In a recent study (Cruse, I., and Maines, M.D. (1988) J. Biol. Chem. 263, 3348-3353), we reported the isolation of small cDNA fragment encoding portion heme oxygenase-2 through immunological screening rat testis library in lambda gt11. We have now used this 274-base pair (bp) as hybridization probe for rescreening same library, thereby recovered number additional positive isolates. Of these, three candidates approximately 900, 1100, 1300 bp, respectively, were subsequently subcloned sequenced. Although differing length, sequences these clones found to be otherwise identical. Moreover, length isolate 18B, 1284 corresponded well with that single mRNA species (approximately 1300-1350 nucleotides) detected Northern blot analysis total poly(A)+RNA. This full- or near full-length encodes 315-amino acid protein molecular weight 35,757, good agreement 36,000 estimated oxygenase-2. When expressed Escherichia coli, cross-reacts antiserum (as assayed by Western immunoblotting) yields high levels oxygenase activity bacterial soluble cell extracts. Finally, computer sequence indicates predicted amino hydropathy profile exhibit similarity oxygenase-1.