Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice
Transgenic mice have been generated that express the E. coli beta-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from â664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of...
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| Published in | Development (Cambridge) Vol. 105; no. 4; pp. 707 - 714 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
England
The Company of Biologists Limited
01.04.1989
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| Subjects | |
| Online Access | Get full text |
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| Abstract | Transgenic mice have been generated that express the E. coli beta-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from â664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the beta-galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. No constitutive expression was observed by in situ staining or Northern analysis at any stage of development, even in tissues that constitutively express the endogenous hsp68 gene. We conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice. |
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| AbstractList | Transgenic mice have been generated that express the E. coli beta-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from â664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the beta-galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. No constitutive expression was observed by in situ staining or Northern analysis at any stage of development, even in tissues that constitutively express the endogenous hsp68 gene. We conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice. ABSTRACT Transgenic mice have been generated that express the E. coli β-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from -664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the β-galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. No constitutive expression was observed by in situ staining or Northern analysis at any stage of development, even in tissues that constitutively express the endogenous hsp68 gene. We conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice. Transgenic mice have been generated than express the E. coli beta -galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68 . Sequences from -664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the beta -galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. The authors conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice. Transgenic mice have been generated that express the E. coli beta-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from -664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the beta-galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. No constitutive expression was observed by in situ staining or Northern analysis at any stage of development, even in tissues that constitutively express the endogenous hsp68 gene. We conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice. |
| Author | J. Rossant S. Clapoff R. Kothary M.D. Perry L.A. Moran S. Darling |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/2557196$$D View this record in MEDLINE/PubMed |
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| References | MacGregor (2023050311230351700_DEVELOP_105_4_707C22) 1987; 13 Hall (2023050311230351700_DEVELOP_105_4_707C13) 1983; 2 Kozak (2023050311230351700_DEVELOP_105_4_707C17) 1983; 47 Dannenberg (2023050311230351700_DEVELOP_105_4_707C4) 1981 Chappell (2023050311230351700_DEVELOP_105_4_707C3) 1986; 45 Lowe (2023050311230351700_DEVELOP_105_4_707C19) 1983; 3 Bellve (2023050311230351700_DEVELOP_105_4_707C1) 1972; 30 Sorger (2023050311230351700_DEVELOP_105_4_707C35) 1987; 6 Graziosi (2023050311230351700_DEVELOP_105_4_707C9) 1980; 214 Edwards (2023050311230351700_DEVELOP_105_4_707C6) 1986; 6 Haas (2023050311230351700_DEVELOP_105_4_707C11) 1983; 306 Lowe (2023050311230351700_DEVELOP_105_4_707C20) 1984; 81 Sanes (2023050311230351700_DEVELOP_105_4_707C33) 1986; 5 Morange (2023050311230351700_DEVELOP_105_4_707C24) 1984; 4 German (2023050311230351700_DEVELOP_105_4_707C8) 1986; 6 Dudler (2023050311230351700_DEVELOP_105_4_707C5) 1984; 38 Perry (2023050311230351700_DEVELOP_105_4_707C31) 1987; 51 Price (2023050311230351700_DEVELOP_105_4_707C32) 1987; 84 Kothary (2023050311230351700_DEVELOP_105_4_707C16) 1987; 121 Feinberg (2023050311230351700_DEVELOP_105_4_707C7) 1983; 132 Goring (2023050311230351700_DEVELOP_105_4_707C10) 1987; 235 Heikkila (2023050311230351700_DEVELOP_105_4_707C14) 1985; 107 Mirault (2023050311230351700_DEVELOP_105_4_707C23) 1982; 1 Parker (2023050311230351700_DEVELOP_105_4_707C28) 1984; 31 O’Malley (2023050311230351700_DEVELOP_105_4_707C27) 1985; 5 Ungewickell (2023050311230351700_DEVELOP_105_4_707C37) 1985; 4 Lee (2023050311230351700_DEVELOP_105_4_707C18) 1984; 259 Slater (2023050311230351700_DEVELOP_105_4_707C34) 1987; 7 Bienz (2023050311230351700_DEVELOP_105_4_707C2) 1984; 81 Pelham (2023050311230351700_DEVELOP_105_4_707C30) 1982; 1 Wu (2023050311230351700_DEVELOP_105_4_707C39) 1984; 311 Lowe (2023050311230351700_DEVELOP_105_4_707C21) 1986; 261 Wittig (2023050311230351700_DEVELOP_105_4_707C38) 1983; 96 Hahnel (2023050311230351700_DEVELOP_105_4_707C12) 1986; 6 Munro (2023050311230351700_DEVELOP_105_4_707C26) 1986; 46 Muller (2023050311230351700_DEVELOP_105_4_707C25) 1985; 1 Kay (2023050311230351700_DEVELOP_105_4_707C15) 1986; 6 Ting (2023050311230351700_DEVELOP_105_4_707C36) 1987; 55 Pelham (2023050311230351700_DEVELOP_105_4_707C29) 1982; 30 Wu (2023050311230351700_DEVELOP_105_4_707C40) 1986; 83 |
| References_xml | – volume: 51 start-page: 227 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C31 article-title: Isolation of a mouse heat shock gene (hsp68) by recombinational screening publication-title: Gene doi: 10.1016/0378-1119(87)90311-8 contributor: fullname: Perry – volume: 96 start-page: 507 year: 1983 ident: 2023050311230351700_DEVELOP_105_4_707C38 article-title: Heat shock gene expression is regulated during teratocarcinoma cell differentiation and early embryonic development publication-title: Devi Biol doi: 10.1016/0012-1606(83)90187-2 contributor: fullname: Wittig – volume: 261 start-page: 2102 year: 1986 ident: 2023050311230351700_DEVELOP_105_4_707C21 article-title: Molecular cloning and analysis of DNA complementary to three mouse Mr =68, (0) heat shock protein mRNAs publication-title: J. biol. Chem doi: 10.1016/S0021-9258(17)35903-3 contributor: fullname: Lowe – start-page: 375 volume-title: In Methods for Studying Mononuclear Phagocytes year: 1981 ident: 2023050311230351700_DEVELOP_105_4_707C4 article-title: Histochemical stains for macrophages in cell smears and tissue sections:,’j-galactosidase, acid phosphatase, nonspecific esterase, succinic dehydrogenase, and cytochrome oxidase doi: 10.1016/B978-0-12-044220-1.50047-7 contributor: fullname: Dannenberg – volume: 55 start-page: 147 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C36 article-title: The nucleotide sequence encoding the hamster 79 kDa glucose-regulated protein (GRP78) and its conservation between hamster and rat publication-title: Gem: contributor: fullname: Ting – volume: 7 start-page: 1906 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C34 article-title: Transcriptional regulation of a heat shock gene in the yeast Saccharomyas cerevisiae publication-title: Mol. Cell. Biol contributor: fullname: Slater – volume: 31 start-page: 273 year: 1984 ident: 2023050311230351700_DEVELOP_105_4_707C28 article-title: A Drosophila RNA poIII transcription factor binds to the regulatory site of an hsp70 gene publication-title: Cell doi: 10.1016/0092-8674(84)90323-4 contributor: fullname: Parker – volume: 6 start-page: 993 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C35 article-title: Cloning and expression of a gene encoding hsc73, the major hsp70-like protein in unstressed rat cells publication-title: EMBO J doi: 10.1002/j.1460-2075.1987.tb04850.x contributor: fullname: Sorger – volume: 214 start-page: 141 year: 1980 ident: 2023050311230351700_DEVELOP_105_4_707C9 article-title: Variability of response of early Drosophila embryos to heat shock publication-title: J. exp. Zool doi: 10.1002/jez.1402140203 contributor: fullname: Graziosi – volume: 84 start-page: 156 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C32 article-title: Lineage analysis in the vertebrate nervous system by retrovirus-mediated gene transfer publication-title: Proc. natn. Acad. Sci. U.S.A doi: 10.1073/pnas.84.1.156 contributor: fullname: Price – volume: 2 start-page: 101 year: 1983 ident: 2023050311230351700_DEVELOP_105_4_707C13 article-title: Expression and regulation of Eschtirichia coli lacZ gene hybrids in mammalian cells publication-title: J. Mol. Appl. 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Cell. Biol contributor: fullname: Lowe – volume: 81 start-page: 2317 year: 1984 ident: 2023050311230351700_DEVELOP_105_4_707C20 article-title: Proteins related to the major heat shock protein are synthesized in unstressed mouse L cells publication-title: Proc. natn. Acad. Sci. U.S.A doi: 10.1073/pnas.81.8.2317 contributor: fullname: Lowe – volume: 46 start-page: 291 year: 1986 ident: 2023050311230351700_DEVELOP_105_4_707C26 article-title: An hsp70-like protein m the ER: identity with the 78Kd glucose-regulated protein and immunoglobulin heavy chain binding protein publication-title: Cell doi: 10.1016/0092-8674(86)90746-4 contributor: fullname: Munro – volume: 121 start-page: 342 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C16 article-title: Cell-lineage specific expression of the mouse hsp68 gene during embryogenesis publication-title: Devi Boil doi: 10.1016/0012-1606(87)90170-9 contributor: fullname: Kothary – volume: 306 start-page: 387 year: 1983 ident: 2023050311230351700_DEVELOP_105_4_707C11 article-title: Immunoglobulin heavy chain binding protein publication-title: Nature, Lond doi: 10.1038/306387a0 contributor: fullname: Haas – volume: 13 start-page: 253 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C22 article-title: Histochemical staining of clonal mammalian cell lines expressing E. coli, β-galactosidase indicates heterogeneous expression of the bacterial gene publication-title: Som. Cell and Mol. Gen doi: 10.1007/BF01535207 contributor: fullname: MacGregor – volume: 81 start-page: 3138 year: 1984 ident: 2023050311230351700_DEVELOP_105_4_707C2 article-title: Developmental control of the heat-shock response in Xenopus publication-title: Proc. natn. Acad. Sci. U.S.A doi: 10.1073/pnas.81.10.3138 contributor: fullname: Bienz – volume: 38 start-page: 391 year: 1984 ident: 2023050311230351700_DEVELOP_105_4_707C5 article-title: Upstream elements necessary for optimal function of the hsp70 promoter in transformed flies publication-title: Cell doi: 10.1016/0092-8674(84)90494-X contributor: fullname: Dudler – volume: 4 start-page: 730 year: 1984 ident: 2023050311230351700_DEVELOP_105_4_707C24 article-title: Altered expression of heat shock proteins in embryonal carcinoma and mouse early embryonic cells publication-title: Mol. Cell. 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Hypmhermia doi: 10.3109/02656738509029277 contributor: fullname: Muller – volume: 1 start-page: 1473 year: 1982 ident: 2023050311230351700_DEVELOP_105_4_707C30 article-title: A synthetic heat shock promoter element confers heat-inducibility on the herpes simplex virus tk gene publication-title: EMBO J doi: 10.1002/j.1460-2075.1982.tb01340.x contributor: fullname: Pelham – volume: 235 start-page: 456 year: 1987 ident: 2023050311230351700_DEVELOP_105_4_707C10 article-title: In situ detection of,’j-galactosidase in lenses of transgenic mice with a y-crystallinllacZ gene publication-title: Science doi: 10.1126/science.3099390 contributor: fullname: Goring – volume: 6 start-page: 555 year: 1986 ident: 2023050311230351700_DEVELOP_105_4_707C8 article-title: The heat shock response in vivo: Experimental induction during mammalian organogenesis publication-title: Tt!ratogenesis, Carcinogenesis and Mutagenesis doi: 10.1002/tcm.1770060609 contributor: fullname: German – volume: 107 start-page: 483 year: 1985 ident: 2023050311230351700_DEVELOP_105_4_707C14 article-title: Acquisition of the heat shock response and thermotolerance during early development of Xenopus laevis publication-title: Devi Biol doi: 10.1016/0012-1606(85)90329-X contributor: fullname: Heikkila – volume: 5 start-page: 3476 year: 1985 ident: 2023050311230351700_DEVELOP_105_4_707C27 article-title: Constitutively expressed rat mRNA encoding a 70-kilodalton heat shock like protein publication-title: Mol. Cell. Biol contributor: fullname: O’Malley – volume: 259 start-page: 4616 year: 1984 ident: 2023050311230351700_DEVELOP_105_4_707C18 article-title: Biochemical characterization of the 94- and 78-kd glucose-regulated proteins in hamster fibroblasts publication-title: J. biol. Chem doi: 10.1016/S0021-9258(17)43091-2 contributor: fullname: Lee – volume: 1 start-page: 1279 year: 1982 ident: 2023050311230351700_DEVELOP_105_4_707C23 article-title: Regulation of heat shock genes: a DNA sequence upstream of the Drosophila hsp70 gene is essential for their induction in monkey cells publication-title: EMBO J doi: 10.1002/j.1460-2075.1982.tb00025.x contributor: fullname: Mirault – volume: 83 start-page: 629 year: 1986 ident: 2023050311230351700_DEVELOP_105_4_707C40 article-title: Human hsp79 promoter contains at least two distinct regulatory domains publication-title: Proc. natn. Acad. Sci. U.S.A doi: 10.1073/pnas.83.3.629 contributor: fullname: Wu – volume: 30 start-page: 71 year: 1972 ident: 2023050311230351700_DEVELOP_105_4_707C1 article-title: Viabihty and survival of mouse embryos following parental exposures to high temperatures publication-title: J. Reprod. Fertil doi: 10.1530/jrf.0.0300071 contributor: fullname: Bellve – volume: 132 start-page: 6 year: 1983 ident: 2023050311230351700_DEVELOP_105_4_707C7 article-title: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity publication-title: Anal. Biochem doi: 10.1016/0003-2697(83)90418-9 contributor: fullname: Feinberg – volume: 6 start-page: 493 year: 1986 ident: 2023050311230351700_DEVELOP_105_4_707C12 article-title: Express10n of the major heat shock protein (hsp70) family during early mouse development publication-title: Teratogenis, Carcinogenis and Mutagmesis doi: 10.1002/tcm.1770060603 contributor: fullname: Hahnel – volume: 47 start-page: 1 year: 1983 ident: 2023050311230351700_DEVELOP_105_4_707C17 article-title: Comparison of initiation of protein synthesis in procaryotes, eucaryotes and organelles publication-title: Micro. Rev doi: 10.1128/mr.47.1.1-45.1983 contributor: fullname: Kozak – volume: 4 start-page: 3385 year: 1985 ident: 2023050311230351700_DEVELOP_105_4_707C37 article-title: The 70-Kd mammalian heat shock proteins are structurally and functionally related to the uncoating protein that releases clathrin triskelia from coated vesicles publication-title: EMBO J doi: 10.1002/j.1460-2075.1985.tb04094.x contributor: fullname: Ungewickell – volume: 6 start-page: 3134 year: 1986 ident: 2023050311230351700_DEVELOP_105_4_707C15 article-title: Efficient transcription of a Caenorhabditis elegans heat shock gene pair in mouse fibroblasts is dependent on multiple promoter elements which can function bidirectionally publication-title: Mol. Cell. Biol contributor: fullname: Kay |
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| Snippet | Transgenic mice have been generated that express the E. coli beta-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68.... ABSTRACT Transgenic mice have been generated that express the E. coli β-galactosidase gene under the control of the promoter from the mouse heat-shock gene,... Transgenic mice have been generated than express the E. coli beta -galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68 .... |
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| SubjectTerms | Animals Bacterial Proteins beta-Galactosidase - genetics Blastocyst - physiology Blotting, Northern Escherichia coli - genetics Galactosidases - genetics Gene Expression Regulation Lac Operon Mice Mice, Transgenic |
| Title | Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice |
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