title: Molecular cloning of IGλ rearrangements using long-distance inverse PCR (LDI-PCR).
abstract: OBJECTIVE
  Malignant cells of mature B-cell origin show tumor-specific clonal immunoglobulin gene (IG) rearrangements, including V(D)J recombinations, nucleotide mutations, or translocations. Rapid molecular cloning of the breakpoint sequence by long-distance inverse PCR (LDI-PCR) has so far been applied to rearrangements targeted to IGH joining, IGH switch, and IGκ regions. We tended to apply LDI-PCR method for cloning of IGλ rearrangements.
  
  
  METHODS
  To identify which IGλ isotype segment was rearranged, we performed Southern blot analysis using isotype-specific probes. We set inverse primers on the telomeric side of each joining region and amplified rearranged bands detected by Southern blot analysis as corresponding PCR products.
  
  
  RESULTS
  All germline IGλ segments were successfully amplified as expected PCR products. We determined breakpoint sequences of five chromosome translocations involving IGλ locus: three novel t(8;22)(q24;q11), one known t(3;22)(q27;q11), and one partially known t(11;22)(q13;q11). Two of the three t(8;22)(q24;q11) were involved in Jλ with a recombination signal sequence and one of three in the first exon of IGLL5, which lies upstream of Jλ1. Three 8q24 breakpoints were widespread at 132, 260 and 366 kb downstream of MYC locus. The t(3;22)(q27;q11) showed a juxtaposition of Jλ2 and the first intron of BCL6, as previously reported. In t(11;22)(q13;q11), 3'UTR of cyclin D1 fused to the constant region of λ7 with nucleotide mutations. We also amplified four Vλ/Jλ recombination sequences.
  
  
  CONCLUSIONS
  Our method is a useful tool for molecular analysis of genetic events in IGλ.
authors.affiliation: Hematology/Oncology, Wakayama Medical University, Wakayama, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Shimanuki
authors.forename: Takashi
authors.initials: T
authors.lastname: Sonoki
authors.forename: Hiroki
authors.initials: H
authors.lastname: Hosoi
authors.forename: Jyuri
authors.initials: J
authors.lastname: Watanuki
authors.forename: Shogo
authors.initials: S
authors.lastname: Murata
authors.forename: Keiki
authors.initials: K
authors.lastname: Kawakami
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Matsuoka
authors.forename: Nobuyoshi
authors.initials: N
authors.lastname: Hanaoka
authors.forename: Hideki
authors.initials: H
authors.lastname: Nakakuma
mesh_terms: D001483:Base Sequence; D056905:Chromosome Breakpoints; D002880:Chromosomes, Human, Pair 11; D002898:Chromosomes, Human, Pair 8; D003001:Cloning, Molecular / Q000379:methods*; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain; D006801:Humans; D007146:Immunoglobulin lambda-Chains / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D014178:Translocation, Genetic
chemical_list: D007146:Immunoglobulin lambda-Chains