Alu-primed polymerase chain reaction for regional assignment of 110 yeast artificial chromosome clones from the human X chromosome: identification of clones associated with a disease locus.
Over 400 yeast artificial chromosome (YAC) clones were isolated from the human X chromosome, and 110 of these were assigned to regions defined by chromosome translocation and deletion breakpoints. Polymerase chain reaction using Alu primers was applied to YAC clones in order to generate probes, to identify overlapping clones, and to derive "fingerprints" and sequence data directly from total yeast DNA. Several clones were identified in regions of medical interest. One set of three overlapping clones was found to cross a chromosomal translocation implicated in Lowe syndrome. The regional assignment of groups of YAC clones provides initiation points for further attempts to develop large cloned contiguous sequences, as well as material for investigation of regions involved in genetic diseases.
Nelson
D L
DL
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Ballabio
A
A
Victoria
M F
MF
Pieretti
M
M
Bies
R D
RD
Gibbs
R A
RA
Maley
J A
JA
Chinault
A C
AC
Webster
T D
TD
Caskey
C T
CT
D001483:Base Sequence; D015139:Blotting, Southern; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D015252:Deoxyribonucleases, Type II Site-Specific; D015723:Gene Library; D030342:Genetic Diseases, Inborn / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014960:X Chromosome
D015345:Oligonucleotide Probes; C024669:endodeoxyribonuclease AluI; D015252:Deoxyribonucleases, Type II Site-Specific