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. 2001 Sep 3;194(5):557–570. doi: 10.1084/jem.194.5.557

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© 2001 The Rockefeller University Press

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Replacement of the J chain exon 1 with the DTA gene. (A) Disruption of the J chain gene and gene replacement by homologous recombination. Genomic map of J chain locus before and after homologous recombination. Digestion with KpnI gives an endogenous band of 5.6 kb while the recombined locus gives a band of 8.0 kb for JDTAneo and 6.4 kb for JDTA after removal of the neo^r gene. Southern blot analysis of the recombined J chain locus showing wt, heterozygous (+/−) JDTAneo, and heterozygous (+/−) JDTA using an outside probe (not present in the construct used for targeting). E, EcoRI; B, BamHI; S, SacI; P, PstI; K, KpnI; and H, HindIII. (B) Expression analysis by RT-PCR for J chain and DTA transcripts in wt, JDTA heterozygous (JDTA^+/−), and JDTA mice. Primers for J chain were located in exon 1 and in exon 4 of the J chain locus. Primers for DTA transcripts were located in the 3′ end of the DTA gene and in the exon 4 of the J chain locus. Shown is also analysis for HPRT transcripts as a cDNA control.

Replacement of the J chain exon 1 with the DTA gene. (A) Disruption of the J chain gene and gene replacement by homologous recombination. Genomic map of J chain locus before and after homologous recombination. Digestion with KpnI gives an endogenous band of 5.6 kb while the recombined locus gives a band of 8.0 kb for JDTAneo and 6.4 kb for JDTA after removal of the neo^r gene. Southern blot analysis of the recombined J chain locus showing wt, heterozygous (+/−) JDTAneo, and heterozygous (+/−) JDTA using an outside probe (not present in the construct used for targeting). E, EcoRI; B, BamHI; S, SacI; P, PstI; K, KpnI; and H, HindIII. (B) Expression analysis by RT-PCR for J chain and DTA transcripts in wt, JDTA heterozygous (JDTA^+/−), and JDTA mice. Primers for J chain were located in exon 1 and in exon 4 of the J chain locus. Primers for DTA transcripts were located in the 3′ end of the DTA gene and in the exon 4 of the J chain locus. Shown is also analysis for HPRT transcripts as a cDNA control.