High expression of the caprine beta-casein gene in transgenic mice.
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.
Persuy
M A
MA
Laboratoire de Génétique Biochimique, I.N.R.A.-C.R.J., Jouy-en-Josas, France.
Stinnakre
M G
MG
Printz
C
C
Mahe
M F
MF
Mercier
J C
JC
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002364:Caseins / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D007122:Immunoelectrophoresis; D008321:Mammary Glands, Animal / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008892:Milk / Q000032:analysis; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics
D002364:Caseins; D012333:RNA, Messenger; D004247:DNA