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  • 1
    ISSN: 1040-452X
    Keywords: Carbonic anhydrase II ; Insertional genesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have developed six transgenic lines of mice with constructs containing presumptive 5′ regulatory regions of carbonic anhydrase II (CA II). Four of the lines contained 1,100 bases of the 5′ flanking region of the human CA II gene, and two transgenic lines resulted from a construct containing 500 bases of the 5′ flanking region of the mouse CA II gene. Tissue-specific expression of the chloramphenicol acetyltransferase (CAT) gene was not obtained in any of the transgenic lines. One of the transgenic lines was found to have high levels of expression of CAT in cerebellum. This expression persisted through multiple generations and was independent of the parental origin of the transgene. On the assumption that the expression was due to the insertion of the transgene in or near a gene expressed normally in cerebellum, homozygous mice were bred for the transgenic insert to see if a mutation might have been induced. Homozygous mice were found and seemed to be normal in all aspects of their phenotype studied. Thus, in this case, neither the insertion of the gene nor the ectopic expression of CAT seemed to be harmful to the animals.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0192-253X
    Keywords: Mouse ; pre-implantation embryo ; cytochrome c oxidase ; antisense inhibition ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: It had not previously been known whether synthesis of nuclear-encoded mitochondrial subunits occurs in pre-implantation embryos. We have used cytoplasmic injections of antisense RNA transcribed in vitro to study this question. Capped, in vitro transcribed RNA antisense to either cytochrome coxidase subunit IV or VIIc injected into each cell at the two-cell stage markedly inhibited synthesis of adenine nucleotides by the 8- to 16-cell stage, whereas injection of the cognate sense RNAs gave levels similar to those previously published for normal embryos. These results strongly suggest that translation of nuclear-encoded mRNAs for mitochondrial subunits is required during pre-implantation development. It was of additional interest that, not only was ATP decreased, but ADP and AMP as well, with the effect that the charge ratio remained constant. The results also suggest, therefore, that the mechanism by which cells normally regulate their charge ratio, thought to be with adenylate deaminase, is already in place. © 1993Wiley-Liss, Inc.
    Additional Material: 2 Tab.
    Type of Medium: Electronic Resource
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