Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia.
We report the cloning of the chromosome 16 p-arm breakpoint involved in inversion 16(p13;q22) associated with subtype of acute myelomonocytic leukemia (AMML) M4Eo. Inter-Alu polymerase chain reaction (PCR) products from a series of interspecific somatic cell hybrids that contain only small portions of the human chromosome 16 p-arm were generated for use as fluorescent in-situ hybridization (FISH) probes. When applied to patient cells, rapid and unambiguous identification of the inversion resulted. Using FISH analysis, cosmid clones associated with the hybrids were identified that bracketed the p-arm breakpoint. A repeat-free fragment of one of these cosmids (35B11) when used as probe on Southern blots from pulsed-field gels identified rearranged macrorestriction fragments in patient DNA. Yeast artificial chromosomes (YACs) were isolated using sequences derived from cosmids flanking 35B11 in a cosmid contig. Of 4 YACs so identified, 3 were shown by FISH to cross the inversion-16 p-arm breakpoint. Therefore, the breakpoint has been molecularly cloned, and identified as being within these 3 YACs. These clones will facilitate the unraveling of the genetic events associated with inversion-16 and are available tools with immediate clinical application.
Liu
P
P
Department of Hematology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Claxton
D F
DF
Marlton
P
P
Hajra
A
A
Siciliano
J
J
Freedman
M
M
Chandrasekharappa
S C
SC
Yanagisawa
K
K
Stallings
R L
RL
Collins
F S
FS
D001483:Base Sequence; D007446:Chromosome Inversion; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal / Q000648:ultrastructure; D002885:Chromosomes, Human, Pair 16; D003001:Cloning, Molecular; D004260:DNA Repair; D005822:Genetic Vectors; D006801:Humans; D017403:In Situ Hybridization; D015479:Leukemia, Myelomonocytic, Acute / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics
D009838:Oligodeoxyribonucleotides