Physical mapping of yeast artificial chromosomes containing sequences from the human beta-globin gene region.
The recently developed technique for cloning genomic DNA fragments of several hundred kilobases or more into yeast artificial chromosomes (YACs) makes it possible to isolate gene families while preserving their structural integrity. We have analyzed five independent yeast clones identified by PCR screening using oligonucleotides derived from the adult human beta-globin gene. Analysis of the five clones containing YACs by conventional and pulsed-field gel electrophoresis revealed that all of the clones include a YAC with sequences from the adult beta-globin gene as expected. One of the clones contains multiple, unstable YACs. Two other clones carry single YACs in which there are at least two unrelated human genomic inserts. The remaining two clones contain single YACs, 150 and 220 kb in size, that contain the entire beta-globin gene family and flanking regions in a single, structurally intact genomic fragment. These should prove useful in future studies of the regulation of expression of genes in the beta-globin gene cluster.
Department of Physiology, University of California, San Francisco 94143-0444.
K M
KM
Gaensler
M
M
Burmeister
B H
BH
Brownstein
P
P
Taillon-Miller
R M
RM
Myers
D002460:Cell Line; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D016172:DNA Fingerprinting; D015342:DNA Probes; D005796:Genes; D005914:Globins / Q000235:genetics*; D006801:Humans; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013789:Thalassemia / Q000235:genetics
D015342:DNA Probes; D005914:Globins; D004247:DNA