Natalie Mincek, Chemistry

The intent of this project was to reconstruct the cytochrome B gene from the extinct mammoth Mammuthus columbi. This was to be accomplished through amplification by PCR the fragments of the cytochrome B gene in order to determine their respective sequences, after which these DNA sequences would be combined to reconstruct the entire sequence of the gene.

With the aid of the funds received from the Office of Research and Creative Work, this project has progressed although it has not yet been completed. Numerous PCR reactions have been performed which have resulted in the amplification a fragment of the cytochrome B gene that corresponds to the carboxy terminal end of the cytochrome B gene product. This fragment also contains a noncoding region and a partial sequence of the tRNA gene that flanks one end of the cytochrome B gene.

Any mutations occurring in the noncoding region would not have been expressed and therefore would not have been selected against. Hence, it is probable that the frequency of mutations will be greater in this noncoding region than any other coding and more conserved region. Detection of mutations in this noncoding fragment, once sequenced, could prove to be very useful in evaluating the evolutionary relationship of the mammoth with its modem day counterpart-the elephant.

If the carboxy terminal DNA fragment is determined to be authentic, this and two other fragments (obtained before funds were received), will be used to construct more specific primers for DNA amplification. This will allow the remaining parts of the cytochrome B gene to be more easily obtained and sequenced.

Due to the degraded state and the instability of ancient DNA, extracting DNA from an 11,000 year old mammoth bone has been very difficult and the greatest hindrance to this project. As a result one aspect of the project has become to determine the most effective and reliable method of extracting DNA from ancient bone.