Panhandle polymerase chain reaction amplifies MLL genomic translocation breakpoint involving unknown partner gene

被引:28
作者
Felix, CA
Kim, CS
Megonigal, MD
Slater, DJ
Jones, DH
Spinner, NB
Stump, T
Hosler, MR
Nowell, PC
Lange, BJ
Rappaport, EF
机构
[1] CHILDRENS HOSP, DEPT PEDIAT, JOSEPH STOKES JR RES INST, DIV GENET, PHILADELPHIA, PA 19104 USA
[2] UNIV PENN, SCH MED, DEPT PATHOL & LAB MED, PHILADELPHIA, PA 19104 USA
[3] UNIV IOWA, DEPT PEDIAT, IOWA CITY, IA 52242 USA
关键词
D O I
10.1182/blood.V90.12.4679.4679_4679_4686
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
We used a new approach called panhandle polymerase chain reaction (PCR) to clone an MLL genomic translocation breakpoint in a case of acute lymphoblastic leukemia of infancy in which karyotype analysis was technically unsuccessful and did not show the translocation partner. Panhandle PCR amplified known MLL sequence 5' of the breakpoint and 3' sequence from the unknown partner gene from a DNA template with an intrastrand loop schematically shaped like a pan with a handle. The 7-kb panhandle PCR product contained the translocation breakpoint in MLL intron 8. The partner DNA included unique nonrepetitive sequences, Alu and mammalian apparent LTR-retrotransposon (MaLR) repetitive sequences, and a region of homology to expressed sequence tags. MaLR sequences have not been found before near leukemia-associated translocation breakpoints. The nonrepetitive sequences were not homologous to known partner genes of MLL. Screening of somatic cell hybrid and radiation hybrid lines by PCR and fluorescence in situ hybridization analysis of normal metaphase chromosomes mapped the partner DNA to chromosome band 4q21. Reverse transcriptase-PCR identified an MLL-AF-4 chimeric mRNA, indicating that panhandle PCR identified a fusion of MLL with a previously uncharacterized AF-4 intronic sequence. panhandle PCR facilitates cloning translocation breakpoints and identifying unknown partner genes. (C) 1997 by The American Society of Hematology.
引用
收藏
页码:4679 / 4686
页数:8
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