Papers by christiane Printz
HAL (Le Centre pour la Communication Scientifique Directe), 1993
European Journal of Biochemistry, 1992
An 18-kb caprine genomic DNA fragment, comprising the beta-casein transcription unit with about 3... more An 18-kb caprine genomic DNA fragment, comprising the beta-casein transcription unit with about 3-kb 5' and 6-kb 3' flanking regions, was microinjected into fertilized one-cell murine eggs. All nine lines of transgenic mice obtained expressed the transgene in their mammary glands, as demonstrated by Northern blot analysis of mRNA in miscellaneous tissues, and qualitative and quantitative analysis of caprine beta-casein in milk, using SDS/PAGE, Western blotting and rocket immunoelectrophoresis. Two lines produced milk containing up to 21-24 mg of the exogenous protein/ml, a yield which is roughly twice that found in goat milk. The yield reached at least 40 mg/ml in some progeny of crossbred G1 transgenic mice. Thus, the investigated gene appears to be a good candidate for making hybrid constructs that might promote an efficient production of valuable foreign proteins in milk.
Nucleic Acids Research, 1988
Gene, 1995
A 5' truncated caprine (ca) K-casein-encoding gene (~cCas) was fused to the 3' end of a 3' trunca... more A 5' truncated caprine (ca) K-casein-encoding gene (~cCas) was fused to the 3' end of a 3' truncated ca ~Cas. The ~:Cas form comprised the 0.8-kb 3' end of intron 2, the remaining part of the transcription unit containing codons-2 to stop 172, and 0.43 kb of the 3' flanking region. The 3Cas form comprised a 3-kb 5' flanking region and the 5' end of the transcription unit terminating 69 bp dowstream from exon 2 which encodes the 15-amino-acid (aa) signal peptide and the first 2 aa of mature 13Cas. The resulting hybrid gene driven by the ~Cas promoter was expressed in all eight lines of transgenic mice investigated, although at different levels. In two lines, the yield of recombinant (re-) ~cCas was > 3 mg/ml of milk. The stage-and mammary tissue-specific expression was similar to that of endogenous ~Cas. The re-~cCas differed from its goat milk counterpart by the occurrence of four extra aa at the N-terminal end, indicating that the signal peptidase released the [3Cas signal peptide. According to sedimentation analyses of murine milk containing > 3 mg re-~cCas/ml, the latter essentially occurred in micelles. Preliminary comparative assays of the behavior of ca %lCas-KCas and a~lCas-re-KCas mixtures upon incremental addition of Ca 2 + showed that re-~cCas had the capacity to protect ~lCaS against Ca 2 +-induced precipitation in forming stable micelles. In contrast, the expression of a reconstituted ca ~cCas gene was in the range of a few lag re-~cCas/ml of milk in the five transgenic lines investigated.
Gene, 1993
Abstract Bovine α s 2 -casein D (CasD) differs from the common type A by the deletion of a stretc... more Abstract Bovine α s 2 -casein D (CasD) differs from the common type A by the deletion of a stretch of 9 amino acids (aa) starting at a position not precisely known, either at aa 50, 51, or 52. The sequence of cloned PCR-amplified genomic DNA from three homozygous cows, two unrelated females carrying the CasD allele and one carrying the CasA allele, did not reveal any deletion and showed two identical nucleotide (nt) substitutions in the 1.7-kb region of both CasD alleles encompassing codons 43–75 in the cDNA encoding α s 2 -CasA. This strongly suggests that the deleted bovine α s 2 -CasD arises from skipping the 27-nt exon, now identified as exon VIII, which encodes aa 51–59 of α s 2 -CasA. The G→T transversion (allele A→D) affecting the last nt of exon VIII, i.e., the 5' consensus splicing site, might be responsible for the altered splicing of the primary transcript of α s 2 - CasD .
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Papers by christiane Printz