The Journal of Experimental Medicine
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Published online 12 March 2001.
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© The Rockefeller University Press, 0022-1007/2001/3/699/ $5.00
The Journal of Experimental Medicine, Volume 193, Number 6, March 19, 2001 699-712

Original Article

Insertion of Phosphoglycerine Kinase (PGK)-Neo 5' of J{lambda}1 Dramatically Enhances VJ{lambda}1 Rearrangement

Tianhe Suna and Ursula Storba
a Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, Illinois 60637

Correspondence to: Ursula Storb, Department of Molecular Genetics and Cell Biology, University of Chicago, 920 E. 58th St., Chicago, IL 60637. Tel:773-702-4440 Fax:773-702-3172 E-mail:stor{at}midway.uchicago.edu.

Gene-targeted mice were generated with a loxP-neomycin resistance gene (neor) cassette inserted upstream of the J{lambda}1 region and replacement of the glycine 154 codon in the C{lambda}1 gene with a serine codon. This insertion dramatically increases V{lambda}1-J{lambda}1 recombination. J{lambda}1 germline transcription levels in pre-B cells and thymus cells are also greatly increased, apparently due to the strong housekeeping phosphoglycerine kinase (PGK) promoter driving the neo gene. In contrast, deletion of the neo gene causes a significant decrease in VJ{lambda}1 recombination to levels below those in normal mice. This reduction is due to the loxP site left on the chromosome which reduces the J{lambda}1 germline transcription in cis. Thus, the correlation between germline transcription and variable (V), diversity (D), and joining (J) recombination is not just an all or none phenomenon. Rather, the transcription efficiency is directly associated with the recombination efficiency. Furthermore, J{lambda}1 and V{lambda}1 germline transcription itself is not sufficient to lead to VJ recombination in T cells or early pre-B cells. The findings may suggest that in vivo: (a) locus and cell type–specific transactivators direct the immunoglobulin or T cell receptor loci, respectively, to a "recombination factory" in the nucleus, and (b) transcription complexes deliver V(D)J recombinase to the recombination signal sequences.

Key Words: PGK-neo, immunoglobulin rearrangement, loxP, transcription, B cells


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