Detection of gamma quanta due to {pi}{sup 0} and {eta} decays by a modular electromagnetic calorimeter with a neutral trigger under a 40-GeV {pi}{sup -} beam
- Institute of High-Energy Physics, Moscow (Russian Federation); and others
We have previously generated a YAC contig of the SMA region on chromosome 5q13 and initiated construction of a corresponding cosmid contig from these YAC clones and a chromosome 5 cosmid library. In order to screen for candidate genes from this area, a cosmid containing CATT1, a microsatellite marker, in linkage disequilibrium with SMA was used to screen a fetal brain library. The largest cDNA clone (2.2 Kb) isolated from this library, GA1, was mapped back to the original cosmid and then characterized. Screening of a spinal cord library with the GA1 cDNA identified a 700 bp clone. Hybridization to a zoo blot revealed cross-species conservation in pig, sheep, goat and horse. Sequencing of GA1 identified 2221 bp with no long open reading frame or poly A tail. Analysis of this sequence with `grail` showed a small potential coding region of 75 bp coding for 25 amino acids. A database search for homologous sequence using `blast` did not show any significant matches. Furthermore, no introns were identified in genomic subclones when compared to the cDNA sequence by using PCR and various combinations of GA1 sequence-derived primers. Experiments with RT-PCR on DNase-treated total mRNA from adult brain, fibroblasts and liver identified the anticipated GA1 product from fibroblast mRNA only. RACE extension of the cDNA clone revealed that we were dealing with the full-length transcript. Subsequent fine mapping with various combinations of GA1 PCR primers to PAC clones in this region identified 1 copy of a novel 300 bp sequence that has undergone a microduplication yielding several copies of this sequence in a 300 Kb segment of the SMA critical region. PCR-based mutational analysis has not revealed any alteration peculiar to SMA chromosomes. A search for polymorphisms with the GA1 sequence to use in SMA linkage analysis was also conducted.
- OSTI ID:
- 133138
- Journal Information:
- Instruments and Experimental Techniques, Vol. 37, Issue 3; Other Information: PBD: Nov 1994; TN: Translated from Pribory i Tekhnika Eksperimenta; No. 3, 43-48(May-Jun 1994)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
BASIC STUDIES
GENES
DNA-CLONING
GENETIC MAPPING
TRANSCRIPTION
DNA SEQUENCING
DISEASES
MUSCLES
HUMAN CHROMOSOME 5
HUMAN POPULATIONS
GENETICS
COMPARATIVE EVALUATIONS
MAMMALS
PATIENTS
YEASTS
CONTIGS
BIOLOGICAL MARKERS
STATISTICS
INTRONS
AMINO ACIDS
POLYMERASE CHAIN REACTION
DNA HYBRIDIZATION