High-level expression of bovine alpha s1-casein in milk of transgenic mice.
The bovine alpha s1-casein gene, isolated from a cosmid library, was introduced into the murine germline. Transgene expression occurred in all transgenic mice, and was confined to the lactating mammary gland. Half of the mouse lines (five out of ten) expressed at relatively high expression levels (> 1 mg ml-1). The highest levels of expression were obtained with a transgene containing 14.2 kb of 5' flanking sequence, in two cases expression levels comparable to (10 mg ml-1) or well above (20 mg ml-1) alpha s1-casein levels in bovine milk were obtained. Transcription initiation occurred at the same site in the bovine alpha s1-casein gene in transgenic mouse as in the cow. A marked induction of expression occurred at parturition rather than at mid-pregnancy, and thus resembled the bovine rather than the murine developmental expression pattern. Bovine alpha s1-casein specific immunoblotting and RIA were developed for characterization and quantification of the recombinant protein. Using these assays, the properties of the recombinant protein could not be distinguished from those of the natural bovine protein. In spite of the high-level tissue-specific and correctly regulated developmental expression of the transgene, expression levels were integration-site dependent. This may indicate that not all cis-acting regulatory elements involved in bovine alpha s1-casein expression were included in the transgene.
Rijnkels
M
M
Medical Biotechnology Dept, Leiden Institute of Chemistry, Leiden University, Netherlands.
Kooiman
P M
PM
Platenburg
G J
GJ
van Dixhoorn
M
M
Nuijens
J H
JH
de Boer
H A
HA
Pieper
F R
FR
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002364:Caseins / Q000096:biosynthesis* / Q000235:genetics*; D002417:Cattle; D003360:Cosmids / Q000235:genetics; D005260:Female; D015870:Gene Expression; D015151:Immunoblotting; D008321:Mammary Glands, Animal / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008892:Milk / Q000378:metabolism*; D011247:Pregnancy; D011863:Radioimmunoassay; D014158:Transcription, Genetic; D019076:Transgenes
D002364:Caseins