Characterization of the K-ras gene of the marsupial Monodelphis domestica.

A cDNA clone isolated from a plasmid library contains the complete coding sequence for the K-ras gene of the marsupial Monodelphis domestica, a South American opossum. The nucleotide sequence of the coding region of the opossum K-ras gene is very similar to the K-ras coding sequences of placental mammals. The coding region of the opossum gene is 95% identical to the human gene at the nucleotide level; the human and opossum genes are 99% identical at the level of encoded amino acids. Transcribed but untranslated regions of the opossum gene 3' and 5' to the coding region are similar to corresponding regions of the human and mouse genes, but are less highly conserved than translated sequences. Based on the nucleotide sequence of the opossum K-ras cDNA clone, primers were designed that allowed amplification of exons 1 and 2 of the gene from opossum genomic DNA by the polymerase chain reaction. When exons 1 and 2 of K-ras were amplified from DNA isolated from an ultraviolet radiation-induced eye tumor of M. domestica and the nucleotide sequence of amplified material was determined, a heterozygous mutation in codon 61 of the gene was detected. This T to A transversion resulted in a change in the amino acid encoded by the codon. The tumor from which DNA was isolated had previously been shown to contain a transforming K-ras oncogene. Thus, the opossum K-ras gene can be mutationally activated in a manner similar to the K-ras genes of placental mammals.