Dominant expression of a 1.3 Mb human Ig kappa locus replacing mouse light chain production

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 Ig kappa 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 Ig kappa disruption, this large human kappa locus replaced the endogenous locus, and subsequent down-regulation of Ig lambda light chain contribution Zed 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 lambda/kappa ratio in serum appears to be the result of competition for early surface Ig expression maintained in the developing B cell.