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Consecutive inactivation of both alleles of the pim-1 proto-oncogene by homologous recombination in embryonic stem cells 总被引:39,自引:0,他引:39
Specific genes can be inactivated or mutated in the mouse germ line. The phenotypic consequences of the mutation can provide pivotal information on the function of the gene in development and maintenance of the mammalian organism. The procedure entails homologous recombination in embryonic stem cells, which, on fusion to recipient blastocysts, give rise to chimaeric mice that can transmit the mutant gene to their offspring. Inbreeding can then yield mice carrying the mutation in both alleles allowing the phenotypic analysis of recessive mutations. In addition to mice lacking a particular gene function, cell lines carrying null alleles of normally expressed genes can be instrumental in assessing the function of the gene. These cell lines can either be obtained from homozygous animals or, should the mutation be lethal early in embryonic development, be generated by consecutive inactivation of both alleles by homologous recombination in cultured cells. Here we illustrate the feasibility of this latter approach by the efficient consecutive inactivation of both alleles of the pim-1 proto-oncogene in embryonic stem cells. 相似文献
174.
Requirement for a functional Rb-1 gene in murine development. 总被引:86,自引:0,他引:86
A R Clarke E R Maandag M van Roon N M van der Lugt M van der Valk M L Hooper A Berns H te Riele 《Nature》1992,359(6393):328-330
Human retinoblastomas can occur both as hereditary and as sporadic cases. Knudson's proposal that they result from two mutational events, of which one is present in the germ line in hereditary cases, has been confirmed by more recent molecular analysis, which has shown both events to involve loss or mutational inactivation of the same gene, RB-1 (ref. 2). RB-1 heterozygosity also predisposes to osteosarcoma, and RB-1 allele losses are seen in sporadic lung, breast, prostate and bladder carcinomas. RB-1 is expressed in most, if not all, tissues and codes for a nuclear phosphoprotein which becomes hypophosphorylated in the G0 growth arrest state and in the G1 phase of the cell cycle. To gain a further insight into the role of RB-1 we and other groups have generated mice carrying an inactivated allele of the homologous gene, Rb-1 (ref. 10), by gene targeting. We report here that young heterozygous mice do not appear abnormal and do not develop retinoblastoma at a detectable frequency. However, homozygous mutant embryos fail to reach term and show a number of abnormalities in neural and haematopoietic development. Broadly similar results are reported by the other groups. 相似文献
175.
Location of high affinity Ca2+-binding sites within the predicted transmembrane domain of the sarcoplasmic reticulum Ca2+-ATPase 总被引:11,自引:0,他引:11
Cation pumps bind and translocate ions with the intermediate formation of a phosphoenzyme. In spite of extensive knowledge of the primary and even secondary structures of several of these cation transport enzymes, however, no high affinity cation binding sites have yet been determined. Here we report the use of oligonucleotide-directed, site-specific mutagenesis to identify the amino acids involved in Ca2+ binding in one of these transport enzymes, the Ca2+-ATPase of sarcoplasmic reticulum. Alteration of Glu 309, Glu 771, Asn 796, Thr 799, Asp 800 or Glu 908, each of which is predicted to lie near the centre of the transmembrane domain in putative transmembrane sequences M4, M5, M6 and M8 resulted in complete loss of Ca2+ transport function and of Ca2+-dependent phosphorylation of the enzyme by ATP. Phosphorylation of each of the mutant enzymes with inorganic phosphate was observed, however, even in the presence of Ca2+, which inhibits phosphorylation in the wild-type enzyme possessing an intact high affinity Ca2+-binding site. These results suggest that at least six polar, oxygen-containing residues lying near the centre of the transmembrane domain provide ligands for one or both of the two high affinity Ca2+ binding sites in the Ca2+-ATPase. 相似文献
176.
L. J. Old D. A. Clarke B. Benacerraf Elisabeth Stockert 《Cellular and molecular life sciences : CMLS》1962,18(7):335-336
Zusammenfassung Die Regression des Sarkomas 180 in Albinomäusen wurde durch vorangehende Splenektomie gefördert. Die gegen Sarkoma 180 erhöhte Resistenz von Mäusen, die mit B.C.G. infiziert worden waren, wird durch die Abwesenheit der Milz nicht beeinflusst.
This work was supported by the American Cancer Society, Inc., New York, N. Y., and The Health Research Council of the City of New York under Contract I-138. 相似文献
This work was supported by the American Cancer Society, Inc., New York, N. Y., and The Health Research Council of the City of New York under Contract I-138. 相似文献
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C. H. Clarke 《Cellular and molecular life sciences : CMLS》1965,21(10):582-583
Zusammenfassung Zehn der vonLeupold isolierten UV-induzierten Mutanten beim Adenin-1-Gen vonSchizosaccharomyces pombe sind in zehn verschiedenen Stellen im Gen lokalisiert.
I am extremely grateful to ProfessorU. Leupold for his help and hospitality in his laboratory, and to Dr.Charlotte Auerbach for her encouragement and interest. 相似文献
I am extremely grateful to ProfessorU. Leupold for his help and hospitality in his laboratory, and to Dr.Charlotte Auerbach for her encouragement and interest. 相似文献