RAG-2 expression is not essential for chicken immunoglobulin gene conversion.
Chicken B cells diversify their immunoglobulin genes by gene conversion in the bursa of Fabricius. The avian leukosis virus-induced B-cell line DT40 continues to diversify its immunoglobulin light chain locus by gene conversion during in vitro passage. Since a variable(diversity)joining recombination-activating gene, RAG-2, is specifically expressed in chicken B cells undergoing immunoglobulin gene conversion, it has been suggested that RAG-2 may be involved in the immunoglobulin gene conversion process. We previously reported high ratios of targeted to random integration after transfection of genomic DNA constructs into DT40. This allows us to easily investigate the function of a gene product by gene disruption. We show here that subclones of DT40 maintain the ability to diversify their immunoglobulin light chain locus by gene conversion even after both copies of the RAG-2 coding regions are deleted. These results demonstrate that the RAG-2 product is not required for gene conversion activity in the immunoglobulin light chain locus.
Takeda
S
S
Basel Institute for Immunology, Switzerland.
Masteller
E L
EL
Thompson
C B
CB
Buerstedde
J M
JM
D000818:Animals; D000916:Antibody Diversity; D001402:B-Lymphocytes / Q000502:physiology; D001483:Base Sequence; D002645:Chickens / Q000235:genetics; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins; D005785:Gene Conversion; D005803:Genes, Immunoglobulin; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007075:Immunoglobulin M / Q000378:metabolism; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics*; D011544:Pseudogenes
D004268:DNA-Binding Proteins; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M; D011506:Proteins; C064655:V(D)J recombination activating protein 2