Isolation of conserved sequences from yeast artificial chromosomes by exon amplification.
Exon amplification is a technique designed to solve a central problem in mammalian molecular genetics--specifically, the isolation of genes from large regions of genomic DNA. This technique allows exons to be isolated from genomic DNA following the selective removal of introns by the eukaryotic splicing mechanism. It is a relatively rapid procedure and can theoretically be applied to test the coding potential of very long stretches of genomic DNA. In this paper we describe the application of exon amplification to a mouse yeast artificial chromosome (YAC) 175 kb in length. A number of potential coding sequences were amplified, and two sequences were shown to be conserved across a wide variety of species representing potential genes.
Gibson
F
F
St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London, UK.
Lehrach
H
H
Buckler
A J
AJ
Brown
S D
SD
North
M A
MA
D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D018244:Chromosomes, Artificial, Yeast; D003001:Cloning, Molecular; D017124:Conserved Sequence; D015245:Deoxyribonuclease BamHI; D015252:Deoxyribonucleases, Type II Site-Specific; D005091:Exons; D015696:Gene Products, tat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D054322:tat Gene Products, Human Immunodeficiency Virus
D001426:Bacterial Proteins; D015696:Gene Products, tat; D054322:tat Gene Products, Human Immunodeficiency Virus; D015245:Deoxyribonuclease BamHI; C024404:BglII endonuclease; D015252:Deoxyribonucleases, Type II Site-Specific