EXPRESSION ANALYSIS OF THE INDIVIDUAL BOVINE BETA-CASEIN, ALPHA(S2)-CASEIN AND KAPPA-CASEIN GENES IN TRANSGENIC MICE

被引:49
作者
RIJNKELS, M
KOOIMAN, PM
KRIMPENFORT, PJA
DEBOER, HA
PIEPER, FR
机构
[1] GENEPHARMING EUROPE BV,2333 CA LEIDEN,NETHERLANDS
[2] LEIDEN UNIV,LEIDEN INST CHEM,DEPT MED BIOTECHNOL,2300 RA LEIDEN,NETHERLANDS
关键词
D O I
10.1042/bj3110929
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
To identify cis-acting regulatory elements involved in the regulation of expression of the casein genes, the bovine beta-, alpha(s2)- and kappa-casein genes were isolated from cosmid libraries and introduced into the murine germline. Bovine casein expression was analysed at the RNA and protein level. The bovine beta-casein gene, including 16 kb of 5 '- and 8 kb of 3 '-flanking region, appeared to be expressed in all 12 transgenic mouse lines analysed. In 50 % of these lines expression levels in milk exceeded 1 mg/ml. Three lines displayed expression levels comparable with or well above (20 mg/ml) the beta-casein levels in bovine milk. Transgene expression was restricted to the mammary gland. Strong induction of expression occurred at parturition and thus resembled the bovine rather than the murine pattern. In spite of this high-level tissue-specific and developmentally regulated expression, beta-casein expression levels were integration-site-dependent, suggesting that not all elements involved in regulation of expression were included in this beta-casein clone. Neither the bovine alpha(s2)- nor the kappa-casein gene, including 8 kb and 5 kb of 5 '- and 1.5 kb and 19 kb of 3 '-flanking sequences respectively, were properly expressed in transgenic mice. However, they were transcribed in stably transfected mouse mammary epithelial cells. This indicates that regulatory elements required for high level, mammary gland-specific expression are not present in the alpha(s2)-and kappa-casein clones used in this study and are probably located elsewhere in the casein gene locus.
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页码:929 / 937
页数:9
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