Nucleic Acids Research, Vol 26, Issue 4 911-918, Copyright © 1998 by Oxford University Press
M Sugai, S Kondo, A Shimizu and T Honjo
Immunoglobulin class switch recombination enables B lymphocytes to
sequentially express antibodies that have identical specificities but that
differ in class and effector function. Although several cis elements
required for class switch recombination have been identified, few trans
-acting factors which are directly related to class switching have been
found. Previously we have developed an efficient in vitro class switching
system using a cell line, CH12F3-2. To clarify the molecular mechanism of
class switching, we intended to isolate genes induced with interleukin
(IL)-4, transforming growth factor (TGF)-beta and CD40L using the in vitro
class switching system. For that purpose, an improved method for making
subtracted cDNA libraries, using uracil DNA glycosylase, has been
developed. This method can overcome a general problem of subtraction, that
rare cDNAs are easily lost. This new subtraction method was applied to the
CH12F3-2 switching system to isolate genes induced by stimulations with
IL-4, TGF-beta and CD40L, and cDNAs encoding deiodinase 1 and SS32, an
alternatively spliced form of the muscle LIM protein, were obtained. Their
expression patterns in response to various combinations of stimuli and the
time courses of the induction supported the usefulness of this method.
ARTICLES
Isolation of differentially expressed genes upon immunoglobulin class switching by a subtractive hybridization method using uracil DNA glycosylase
Department of Medical Chemistry, Faculty of Medicine and Center for Molecular Biology and Genetics, Kyoto University Sakyo-ku, Kyoto 606, Japan.
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