| First Author | Gonzalez FJ | Year | 1981 |
| Journal | J Biol Chem | Volume | 256 |
| Issue | 10 | Pages | 4697-700 |
| PubMed ID | 6262298 | Mgi Jnum | J:6506 |
| Mgi Id | MGI:54982 | Doi | 10.1016/s0021-9258(19)69304-x |
| Citation | Gonzalez FJ, et al. (1981) Cloning of epoxide hydratase complementary DNA. J Biol Chem 256(10):4697-700 |
| abstractText | Tightly membrane-bound polysomes were isolated from livers of rats administered trans-stilbene oxide. Epoxide hydratase mRNA was enriched from these polysomes using immunochemical techniques and oligo(dT)-cellulose chromatography. This resulted in an increase in message concentration over that found in noninduced membrane-bound cDNA, synthesized from enriched mRNA, was inserted into the ampicillin resistance gene of pBR322 using oligo(dG)-oligo(dC) tailing. Clones containing sequences complementary to epoxide hydratase mRNA were selected by differential colony hybridization using [32P]cDNA synthesized from immunoenriched mRNA and [32P]cDNA synthesized from nonenriched mRNA. Plasmids from four clones, which only annealed with the enriched probe, were isolated and shown to specifically hybridize with epoxide hydratase mRNA by hybrid selection-translation. A composite restriction endonuclease map of the plasmid inserts was constructed which spanned 1310 base pairs and represented approximately 80% of the message sequence. The 3'-5' orientation of this map relative to the epoxide hydratase mRNA was also determined. |
