Meredith McCoy Cloning Methionine Aminopeptidase from the Extreme Halophile Haloarcula marismortui Division Sciences Department Biology Year 2003 Advisor Charles Sokolik

Haloarcula marismortui is an archaebacteria that thrives in a 6 M saline environment.  To compensate for the osmotic pressure from its environment, H. marismortui accumulates K+ and other ions intracellularly, which necessitates the adaptation of its proteins to function in high salt environments.  Methionine aminopeptidase (MAP), a ubiquitous protein responsible for cleaving the initial methionine of many proteins as a common post-translational modification, will be cloned from H. marismortui and its adaptations to a high salt environment investigated.  The gene for MAP from H. marismortui (HmMAP) will be cloned using the polymerase chain reaction (PCR).  We present here the appropriate primers and the optimal conditions for the PCR amplification of HmMAP, as well as the sequence of a successfully cloned segment of the gene.  The entire HmMAP gene could be located by using this segment of the gene as a probe in Southern hybridizations.