Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia

P. Liu, D. F. Claxton, P. Marlton, A. Hajra, J. Siciliano, M. Freedman, S. C. Chandrasekharappa, K. Yanagisawa, R. L. Stallings, F. S. Collins, M. J. Siciliano

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)716-721
Number of pages6
JournalBlood
Volume82
Issue number3
StatePublished - Jan 1 1993

Fingerprint

Leukemia, Myelomonocytic, Acute
Yeast Artificial Chromosomes
Cosmids
Chromosomes
Yeast
Fluorescence In Situ Hybridization
Chromosomes, Human, Pair 16
Clone Cells
Hybrid Cells
Human Chromosomes
Southern Blotting
Cloning
Polymerase chain reaction
Organism Cloning
Reaction products
Gels
Polymerase Chain Reaction
DNA

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Liu, P., Claxton, D. F., Marlton, P., Hajra, A., Siciliano, J., Freedman, M., ... Siciliano, M. J. (1993). Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia. Blood, 82(3), 716-721.
Liu, P. ; Claxton, D. F. ; Marlton, P. ; Hajra, A. ; Siciliano, J. ; Freedman, M. ; Chandrasekharappa, S. C. ; Yanagisawa, K. ; Stallings, R. L. ; Collins, F. S. ; Siciliano, M. J. / Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia. In: Blood. 1993 ; Vol. 82, No. 3. pp. 716-721.
@article{793edfee80fa4455bfff9998b2246b5f,
title = "Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia",
abstract = "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.",
author = "P. Liu and Claxton, {D. F.} and P. Marlton and A. Hajra and J. Siciliano and M. Freedman and Chandrasekharappa, {S. C.} and K. Yanagisawa and Stallings, {R. L.} and Collins, {F. S.} and Siciliano, {M. J.}",
year = "1993",
month = "1",
day = "1",
language = "English (US)",
volume = "82",
pages = "716--721",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "3",

}

Liu, P, Claxton, DF, Marlton, P, Hajra, A, Siciliano, J, Freedman, M, Chandrasekharappa, SC, Yanagisawa, K, Stallings, RL, Collins, FS & Siciliano, MJ 1993, 'Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia', Blood, vol. 82, no. 3, pp. 716-721.

Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia. / Liu, P.; Claxton, D. F.; Marlton, P.; Hajra, A.; Siciliano, J.; Freedman, M.; Chandrasekharappa, S. C.; Yanagisawa, K.; Stallings, R. L.; Collins, F. S.; Siciliano, M. J.

In: Blood, Vol. 82, No. 3, 01.01.1993, p. 716-721.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of yeast artificial chromosomes containing the inversion 16 p-arm breakpoint associated with acute myelomonocytic leukemia

AU - Liu, P.

AU - Claxton, D. F.

AU - Marlton, P.

AU - Hajra, A.

AU - Siciliano, J.

AU - Freedman, M.

AU - Chandrasekharappa, S. C.

AU - Yanagisawa, K.

AU - Stallings, R. L.

AU - Collins, F. S.

AU - Siciliano, M. J.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0027304591&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027304591&partnerID=8YFLogxK

M3 - Article

C2 - 8338941

AN - SCOPUS:0027304591

VL - 82

SP - 716

EP - 721

JO - Blood

JF - Blood

SN - 0006-4971

IS - 3

ER -