Nucleic Acids Research, Vol 25, Issue 15 3066-3073, Copyright © 1997 by Oxford University Press
J Du, Y Zhu, A Shanmugam and AL Kenter
The molecular mechanism of immunoglobulin switch recombination is poorly
understood. Switch recombination occurs between pairs of switch regions
located upstream of the constant heavy chain genes. Previously we showed
that switch recombination breakpoints cluster to a defined subregion in the
Sgamma3, Sgamma1 and Sgamma2b tandem repeats. We have developed a strategy
for direct amplification of Smu/Sgamma3 composite fragments as well as Smu
and Sgamma3 regions by PCR. This assay has been used to analyze the
organization of Smu, Sgamma3 and a series of Smu/Sgamma3 recombination
breakpoints from hybridomas and normal mitogen-activated splenic B cells.
DNA sequence analysis of the switch fragments showed direct joining of Smu
and Sgamma3 without deletions or duplications. Mutations were found in two
switch junctions on both sides of the crossover point, suggesting that
template switching is the most likely model for the mechanism of switch
recombination. Statistical analysis of the positions of the recombination
breakpoints in the Sgamma3 tandem repeat indicates the presence of two sub-
clusters, suggesting non-random usage of DNA substrate in the recombination
reaction.
ARTICLES
Analysis of immunoglobulin Sgamma3 recombination breakpoints by PCR: implications for the mechanism of isotype switching
Department of Microbiology and Immunology (M/C 790), University of Illinois College of Medicine, 835 South Wolcott Avenue, Chicago, IL 60612-7344, USA.
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