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Journal of Neuroscience, Vol 14, 2290-2300, Copyright © 1994 by Society for Neuroscience
Cloning and characterization of the choline acetyltransferase structural gene (cha-1) from C. elegans
A Alfonso, K Grundahl, JR McManus and JB Rand
Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, Oklahoma City 73104.
We have cloned the cha-1 gene from Caenorhabditis elegans using the method
of transposon tagging, cha-1 is the structural gene for ChAT, the enzyme
that synthesizes ACh. Sequence analysis of cDNAs predicts a protein of 71.5
kDa; comparison of the deduced amino acid sequence with ChAT sequences from
other species confirms that cha-1 encodes ChAT. Comparison of cDNA and
genomic sequences reveals that transcription is from right to left on the
genetic map, and that some of the transcripts may result from
trans-splicing of the 22-base spliced leader SL 1. The cha-1 gene is
organized into 11 exons. The first exon contains only untranslated
sequences, and is followed by an extremely long intron. The coding sequence
of the cha-1 transcript is disrupted by mutations in the cha-1 gene. We
have determined the sites of four transposon insertions and the end-points
of two deletions that lead to the cha-1 mutant phenotype; one of the
deletions appears to eliminate gene function completely. Comparison of the
Drosophila, rat, and C. elegans genes reveals conserved motifs and
conserved intron sites.
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