Structure of the rabbit tc-casein encoding gene: expression of the cloned gene in the mammary gland of transgenic mice

The rabbit κ-casein (κ-Cas) encoding gene has been isolated as a series of overlapping DNA fragments cloned from a rabbit genomic library constructed in bacteriophage λEMBL3. The clones harboured the 7.5-kb gene flanked by about 2.1 kb upstream and 9 kb downstream sequences. The cloned gene is the most frequently occurring of two κ-Cas alleles identified in New Zealand rabbits. Comparison of the corresponding domains in rabbit and bovine κ-Cas shows that both genes comprise 5 exons and that the exon/intron boundary positions are conserved whereas the introns have diverged considerably. The first three introns are shorter in the rabbit, the second intron showing the greatest difference between the two species: 1.35 kb instead of 5.8 kb in the bovine gene. Repetitive sequence motives reminiscent of the rabbit C type repeat and the complementary inverted C type repeat were identified in the fourth and first introns, respectively. Transgenic mice were produced by microinjecting into mouse oocytes an isolated genomic DNA fragment which contained the entire κ-Cas coding region, together with 2.1-kb 5′ and 4.0-kb 3′ flanking region. Expression of transgene rabbit κ-Cas mRNA could be detected in the mammary gland of lactating transgenic mice and the production of rabbit κ-Cas was detected in milk using species-specific antibodies. The cloned gene is thus functional.