Abstract
We have developed a simple, straightforward procedure to isolate exons from cloned human genomic DNA. The method is PCR based and relies upon the conservation of splice-site sequences and the frequency of Alu repeat elements in the genome to capture coding sequences. We designed two different sets of primers: a primer from each end of the Alu element and primers with the 5' or 3' splice-site consensus sequences. Putative exons were amplified by PCR using YAC DNA as starting material. We applied Alu-splice PCR to two overlapping YACs, 72H9 and 860G11, from human chromosome 21. Sequence and northern analysis of 37 initial clones resulted in the identification of five novel exons.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Amino Acid Sequence
-
Base Sequence
-
Chromosome Mapping
-
Chromosomes, Artificial, Yeast
-
Chromosomes, Human, Pair 21 / genetics*
-
Cloning, Molecular / methods*
-
DNA Primers
-
DNA-Binding Proteins
-
Down Syndrome / genetics
-
Exons*
-
Gene Expression
-
Humans
-
Intracellular Signaling Peptides and Proteins
-
Molecular Sequence Data
-
Muscle Proteins*
-
Pilot Projects
-
Polymerase Chain Reaction / methods*
-
Proteins / genetics
-
RNA Splicing
-
Repetitive Sequences, Nucleic Acid*
-
Selection, Genetic
Substances
-
DNA Primers
-
DNA-Binding Proteins
-
Intracellular Signaling Peptides and Proteins
-
Muscle Proteins
-
Proteins
-
RCAN1 protein, human
Associated data
-
GENBANK/U28833
-
GENBANK/U84488
-
GENBANK/U84489
-
GENBANK/U84490
-
GENBANK/U84491
-
GENBANK/U84492
-
GENBANK/U84493
-
GENBANK/U84494