Yeast artificial chromosome cloning of a two-megabase-size contig within chromosomal band 18q21 establishes physical linkage between BCL2 and plasminogen activator inhibitor type-2.
The construction of large-scale physical maps requires efficient approaches to generate new probes and link informative markers. The mapping of a human chromosomal segment was initiated by using the 18q21.3 probes, plasminogen activator inhibitor type-2 (PLANH2) and BCL2, to screen a yeast artificial chromosome (YAC) library. An inverse polymerase chain reaction technique rescued genomic ends of the YAC inserts. These new probes were used in a chromosomal walking strategy which established that the PLANH2 gene was 600 kb telomeric and in the opposite transcriptional orientation to that of BCL2. Overall, 16 YACs with a mean size of approximately 300 kb were analyzed using rare-cutting restriction endonucleases and 10 end-rescued probes. A contiguous map within 18q21.3 that spans approximately 2 Mb was assembled. This establishes the feasibility of using YACs in the efficient cloning and physical surveying of expanses of the human genome lacking closely spaced, genetic landmarks.
Silverman
G A
GA
Department of Pediatrics, Howard Hughes Medical Institute, St. Louis, Missouri.
Jockel
J I
JI
Domer
P H
PH
Mohr
R M
RM
Taillon-Miller
P
P
Korsmeyer
S J
SJ
D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping / Q000379:methods*; D016386:Chromosome Walking; D015825:Chromosomes, Fungal; D002887:Chromosomes, Human, Pair 18; D003001:Cloning, Molecular; D006224:Cricetinae; D015342:DNA Probes; D015723:Gene Library; D008040:Genetic Linkage; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D015849:Plasminogen Inactivators; D016133:Polymerase Chain Reaction; D011519:Proto-Oncogenes / Q000235:genetics*; D015183:Restriction Mapping; D014158:Transcription, Genetic
D015342:DNA Probes; D015849:Plasminogen Inactivators