Quantification of hprt gene deletions mediated by illegitimate V(D)J recombination in peripheral blood cells of humans
✍ Scribed by James C. Fuscoe; Lisa K. Vira; Deborah D. Collard; Martha M. Moore
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 154 KB
- Volume
- 29
- Category
- Article
- ISSN
- 0893-6692
No coin nor oath required. For personal study only.
✦ Synopsis
V(D)J recombinase is normally involved in the highly nested PCR with two sets of primers. Multiple repliregulated rearrangement of immunoglobulin and T-cates of genomic DNA (each representing 4 1 10 5 cell-receptor gene segments (in B and T cells, re-cells) are amplified with specific primers under conspectively) to form functional antibody genes and ditions in which a single copy will give a detectable T-cell-receptor genes. Occasionally, this tightly con-PCR product. Poisson statistics are then used to estitrolled process acts on inappropriate places in the mate the deletion mutant frequency. The frequency genome and results in deletions and translocations. of cells with the hprt deletion among 20 healthy Some of these illegitimate V(D)J recombinase-medi-young adults ranged from õ1.3 1 10 07 to 4.1 1 ated events have been implicated in the genetic 10 07 and was compared with the frequency of changes associated with several forms of leukemia t(14;18) previously determined in these same indiand lymphoid malignancy. We have developed a viduals. No correlation was found between the fresensitive, specific polymerase chain reaction (PCR)-quencies of these two measures of genomic rearbased assay to quantify such events in the periph-rangement. The DNA sequences at the deletion eral blood cells of humans. This assay detects a junctions were determined and provided evidence V(D)J recombinase-mediated deletion in the hprt for multiple independent mutations in some individugene, which codes for a housekeeping enzyme and als. This assay may serve as a biomarker for the is not implicated in cancer development. Alterations level of illegitimate V(D)J recombination occurring in this gene serve as a surrogate indicator for these in peripheral blood cells of humans. Environ. illegitimate events, which may be occurring Mol. Mutagen. 29:28