Mecr^{tm1(fabI)Akast}

A sequence encoding the bacterial fabI ETR (2-enoyl-thioester reductase), a functional prokaryotic ortholog of Mecr, was introduced into exon 2 of the mouse Mecr gene. The construct was C-terminally tagged with a double HA epitope tag. The splicing acceptor sites of exon 2 and 54 nucleotides of the exon 2 itself were maintained in-frame with the fabI coding region. The Mecr mitochondrial targeting sequence (MTS) is encoded by exon 1. These alterations are predicted to yield a chimeric mRNA encoding HA-tagged fabI N-terminally extended with the Mecr MTS. The construct was followed by the strong transcriptional terminator BGH attached to an FRT-flanked selection cassette to exclude the possibility of producing intact Mecr. (J:82809)