POLYMERASE CHAIN-REACTION AND DNA-SEQUENCE OF RAINBOW-TROUT TUMOR-SUPPRESSOR GENE-P53 EXON-5, EXON-6 AND EXON-7 TO EXON-9

A protocol for the extraction of genomic DNA from teleost fish was dev eloped and produced high molecular weight DNA from English sole, white sucker and rainbow trout. A protocol for polymerase chain reaction (P CR) of tumour suppressor gene p53 from rainbow trout genomic DNA was d eveloped. Several primers were chosen inside exon 5 and 9 and exon 6 a nd 7. A successful PCR method yielded the conserved exon 5 to 9 region of genomic rainbow trout p53 in two fragments: fragment 1 comprising exons 5 and 6 and fragment 2 spanning the region of exons 7 to 9. The apparent molecular weight of fragments 1 and 2 were 510 and 760 base p airs respectively. The sequences of both fragments including the intro ns were determined and gave 515 and 793 bp for the two PCR fragments. These sequences will facilitate examination of tumour suppressor gene p53 conserved exons from genomic DNA e.g. for study of p53 mutations i n fish.