Isolation of candidate genes from the Batten Disease (JNCL) region using exon amplification

Journal Article · Thu Sep 01 04:00:00 EDT 1994 · American Journal of Human Genetics

OSTI ID:134435

Lerner, T J; Haines, J L; Buckler, A ^[1]

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1. Massachusetts General Hospital, Charlestown, MA (United States)

Batten Disease (juvenile neuronal cercoid lipofuscinosis; JNCL) is the most common neurodegenerative disorder of childhood. Clinically, this autosomal recessive disorder is characterized by loss of vision, seizures, and progressive encephalopathy. The JNCL gene CLN3 has been genetically mapped to a 2 cM region in 16p12.1-11.2 and shows significant allelic association with alleles at five marker loci, D16S288, D16S272, D16S299, D16S298, and SPN, within this interval. Extended haplotype analysis strongly suggests that CLN3 must lie in the vicinity of D16S299/D16S298. We have used these markers as STSs to isolate YACs from the CEPH and Los Alamos flow-sorted chromosome 16 YAC libraries. These YACs, in turn, have been used to generate cosmid contigs by hybridization of inter-Alu PCR products to the gridded Los Alamos chr. 16 cosmid library and by direct sub-cloning. In order to identify genes originating from this region we have used the technique of exon amplification. This procedure produces trapped exon clones, which can represent single exons or multiple exons spliced together and is an efficient method for obtaining probes for physical mapping and for screening cDNA libraries. In all experiments, we have analyzed cosmid DNA using a modification of the original exon amplification procedure that increases both the efficiency and sensitivity of the approach. We have used the resulting exon probes to screen retina and brain cDNA libraries for candidate JNCL genes. Our preliminary studies indicate that the Batten candidate region is gene-rich.

• OSTI ID: 134435
• Report Number(s): CONF-941009--
• Journal Information: American Journal of Human Genetics, Journal Name: American Journal of Human Genetics Journal Issue: Suppl.3 Vol. 55; ISSN AJHGAG; ISSN 0002-9297
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
BIOLOGICAL MARKERS
CHILDREN
DNA HYBRIDIZATION
DNA-CLONING
EFFICIENCY
EXONS
GENE AMPLIFICATION
GENES
GENETIC MAPPING
HEREDITARY DISEASES
HUMAN CHROMOSOME 16
NERVOUS SYSTEM DISEASES
POLYMERASE CHAIN REACTION
PROBES
RECESSIVE MUTATIONS
SENSITIVITY
SPLICING
YEASTS