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.
引用
收藏
页码:929 / 937
页数:9
相关论文
共 47 条
[41]   EXTRACELLULAR-MATRIX AND HORMONES TRANSCRIPTIONALLY REGULATE BOVINE BETA-CASEIN 5' SEQUENCES IN STABLY TRANSFECTED MOUSE MAMMARY CELLS [J].
SCHMIDHAUSER, C ;
BISSELL, MJ ;
MYERS, CA ;
CASPERSON, GF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (23) :9118-9122
[42]   A NOVEL TRANSCRIPTIONAL ENHANCER IS INVOLVED IN THE PROLACTIN AND EXTRACELLULAR MATRIX-DEPENDENT REGULATION OF BETA-CASEIN GENE-EXPRESSION [J].
SCHMIDHAUSER, C ;
CASPERSON, GF ;
MYERS, CA ;
SANZO, KT ;
BOLTEN, S ;
BISSELL, MJ .
MOLECULAR BIOLOGY OF THE CELL, 1992, 3 (06) :699-709
[43]   MAMMARY GLAND-SPECIFIC NUCLEAR FACTOR ACTIVITY IS POSITIVELY REGULATED BY LACTOGENIC HORMONES AND NEGATIVELY BY MILK STASIS [J].
SCHMITT-NEY, M ;
HAPP, B ;
HOFER, P ;
HYNES, NE ;
GRONER, B .
MOLECULAR ENDOCRINOLOGY, 1992, 6 (12) :1988-1997
[44]   BETA-CASEIN GENE PROMOTER ACTIVITY IS REGULATED BY THE HORMONE-MEDIATED RELIEF OF TRANSCRIPTIONAL REPRESSION AND A MAMMARY-GLAND-SPECIFIC NUCLEAR FACTOR [J].
SCHMITT-NEY, M ;
DOPPLER, W ;
BALL, RK ;
GRONER, B .
MOLECULAR AND CELLULAR BIOLOGY, 1991, 11 (07) :3745-3755
[45]   MOLECULAR-CLONING OF MOUSE MAMMARY-GLAND KAPPA-CASEIN - COMPARISON WITH RAT KAPPA-CASEIN AND RAT AND HUMAN GAMMA-FIBRINOGEN [J].
THOMPSON, MD ;
DAVE, JR ;
NAKHASI, HL .
DNA-A JOURNAL OF MOLECULAR & CELLULAR BIOLOGY, 1985, 4 (04) :263-271
[46]   GENOMIC ANALYSIS OF THE MAJOR BOVINE-MILK PROTEIN GENES [J].
THREADGILL, DW ;
WOMACK, JE .
NUCLEIC ACIDS RESEARCH, 1990, 18 (23) :6935-6942
[47]   EVOLUTION OF THE CASEIN MULTIGENE FAMILY - CONSERVED SEQUENCES IN THE 5' FLANKING AND EXON REGIONS [J].
YULEE, LY ;
RICHTERMANN, L ;
COUCH, CH ;
STEWART, AF ;
MACKINLAY, AG ;
ROSEN, JM .
NUCLEIC ACIDS RESEARCH, 1986, 14 (04) :1883-1902