Journal article
Drosophila Choline Acetyltransferase Temperature-Sensitive Mutants
Neurochemical research, Vol.24(8), pp.1081-1087
08/1999
DOI: 10.1023/A:1021021213625
PMID: 10478949
Abstract
We used the reverse transcription-polymerase chain reaction (RT-PCR) to amplify choline acetyltransferase (ChAT) mRNA fragments from two temperature-sensitive alleles of Drosophila melanogaster, Cha
ts1 and Cha
ts2. Single base substitutions in the mutants (T1614A in Cha
ts1 and G1596A in Cha
ts2) would result in amino acid changes for ChAT protein (Met403Lys in Cha
ts1 and Arg397His in Cha
ts2). These base substitutions were confirmed in mRNA extracted from homozygous mutants using a Single Nucleotide Primer Extension assay (SNuPE) and are sufficient to produce thermolabile enzyme. Our results indicate that these temperature-sensitive mutants are point mutations in the structural gene for ChAT. Using a quantitative SNuPE assay we also show that similar levels of Cha
ts and wild type transcripts are present in heterozygous flies (Cha
ts1/+ and Cha
ts2
/+) at both restrictive and permissive temperatures. This contrasts with RNase protection assays of ChAT mRNA in homozygous mutant animals where the levels of mutant mRNA decrease at restrictive temperature.
Details
- Title: Subtitle
- Drosophila Choline Acetyltransferase Temperature-Sensitive Mutants
- Creators
- Weiya Wang - Division of Neurosciences Beckman Research Institute of the City of Hope 1450 E. Duarte Rd. Duarte CA 91010Toshihiro Kitamoto - Division of Neurosciences Beckman Research Institute of the City of Hope 1450 E. Duarte Rd. Duarte CA 91010Paul Salvaterra - Division of Neurosciences Beckman Research Institute of the City of Hope 1450 E. Duarte Rd. Duarte CA 91010
- Resource Type
- Journal article
- Publication Details
- Neurochemical research, Vol.24(8), pp.1081-1087
- Publisher
- Kluwer Academic Publishers-Plenum Publishers; New York
- DOI
- 10.1023/A:1021021213625
- PMID
- 10478949
- ISSN
- 0364-3190
- eISSN
- 1573-6903
- Language
- English
- Date published
- 08/1999
- Academic Unit
- Iowa Neuroscience Institute; Anesthesia; Neuroscience and Pharmacology
- Record Identifier
- 9984006322602771
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