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Dominant expression of a 1.3 Mb human IgK locus replacing mouse light chain production
Author(s) -
Zou Xiangang,
Xian Jian,
Davies Nicholas P.,
Popov Andrei V.,
Brüggemann Marianne
Publication year - 1996
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.10.10.8751726
Subject(s) - locus (genetics) , biology , immunoglobulin light chain , gene , cosmid , genetics , germline , antibody repertoire , microbiology and biotechnology , antibody
Expression studies of multigene families, such as the immunoglobulin (Ig) loci, are difficult because of their large size and the necessity to introduce germline configured regions into an animal. Antibody diversity from Ig gene miniloci is limited by the number of variable (V) region genes and the need for distal regulatory elements to control expression. Here, we show germline transfer into mice of a 1300 kb human Igκ light chain locus on a yeast artificial chromosome that resulted in early DNA rearrangement and highly efficient human light chain expression. The human locus was assembled from a 300 kb authentic region using contig extension by addition of cosmid multimers to supplement the variable gene cluster. This resulted in the addition of about 100 V region genes in germline configuration from different families. In transgenic animals with Igκ disruption, this large human κ locus replaced the endogenous locus, and subsequent down‐regulation of Igλ light chain contribution led to a dominant expression of the rearranged human genes. Contrary to expectation, rather than providing a solely selective advantage for ensuring repertoire formation controlled by the sheer number of introduced genes, the λ/κ ratio in serum appears to be the result of competition for early surface Ig expression maintained in the developing B cell.—Zou, X., Xian, J., Davies, N. P., Popov, A. V., Brüggemann, M. Dominant expression of a 1.3 Mb human Igκ locus replacing mouse light chain production. FASEB J. 10, 1227‐1232 (1996)