Reprogrammed expression of subunit 9 of the mitochondrial ATPase complex of Saccharomyces cerevisiae. Expression in vitro from a chemically synthesized gene and import into isolated mitochondria.

摘要: A synthetic gene has been designed and constructed by total chemical synthesis as a first step in the functional relocation from mitochondrion to nucleus of encoding subunit 9 yeast mitochondrial ATPase complex. This (NAP9) incorporates codons frequently used nuclear genes Saccharomyces cerevisiae additionally includes series unique restriction enzyme cleavage sites facilitate future systematic manipulations its protein product. Following expression NAP9 transcription translation vitro, radiolabelled was produced which displayed gel electrophoretic mobility solubility chloroform/methanol characteristic authentic proteolipid encoded vivo olil gene. In order achieve import into mitochondria 9, fusion made between DNA cleavable presequence nuclearly precursor Neurospora crassa. resultant imported appropriately processed isolated mitochondria. The less efficient than that observed parallel experiments with 8 attached same or naturally occurring intact N. crassa precursor. Yeast lacking leader sequence is not but, unlike 8, it does embed itself outer membrane, spite highly hydrophobic character.