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General Comment |
Molecular cloning and characterization of a new gene, Oocyte-G1. Zhang Y et al. Oocytes are recognized as a source of regulatory molecules that influence follicular development through an array of actions on granulosa cells. Recently, more and more hormones and signaling molecules were identified during follicular developmental processes; however, the details about their functions are still unclear. During efforts to clone follicular development-related genes, we isolated a cDNA fragment by DDRT-PCR. To obtain cDNA 5'- and 3'-end sequences, we screened a mouse ovarian cDNA library. After screening the library, an open reading frame of 2,994 bp for the new gene (Oocyte-G1), which encodes a 997-residue protein, was cloned. Northern blot analysis revealed the presence of approximately 3.6 kb Oocyte-G1 mRNA in ovary, lung, kidney, testis and brain. Northern analysis of RNA from ovaries in vivo showed that Oocyte-G1 was weakly expressed on day 5 and at a moderate level on day 10. Thereafter, on day 15 or in adults (day 40), there was an increase in expression, followed by a decline in ovaries on day 20 or older (day 120). Furthermore, we studied the Ooctye-G1 protein by using the antiserum against a peptide sequence unique to this gene in Western blotting and immunolocalization. The antiserum recognized a prominent band of approximately 110 kDa in immunoblots and signals were dispersed in oocytes and some cumulus granulosa cells. Our results suggest the potential role of Oocyte-G1 in ovarian follicular development. J. Cell. Physiol. (c) 2008 Wiley-Liss, Inc.
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Mutations |
1 mutations
Species: mouse
Mutation name: None
type: targeted overexpression
fertility: subfertile
Comment: Production of transgenic mice by random recombination of targeted genes in female germline stem cells. Zhang Y et al. Oocyte production in most mammalian species is believed to cease before birth. However, this idea has been challenged with the finding that postnatal mouse ovaries possess mitotically active germ cells. A recent study showed that female germline stem cells (FGSCs) from adult mice were isolated, cultured long term and produced oocytes and progeny after transplantation into infertile mice. Here, we demonstrate the successful generation of transgenic or gene knock-down mice using FGSCs. The FGSCs from ovaries of 5-day-old and adult mice were isolated and either infected with recombinant viruses carrying green fluorescent protein, Oocyte-G1 or the mouse dynein axonemal intermediate chain 2 gene, or transfected with the Oocyte-G1 specific shRNA expression vector (pRS shOocyte-G1 vector), and then transplanted into infertile mice. Transplanted cells in the ovaries underwent oogenesis and produced heterozygous offspring after mating with wild-type male mice. The offspring were genetically characterized and the biological functions of the transferred or knock-down genes were investigated. Efficiency of gene-transfer or gene knock-down was 29%-37% and it took 2 months to produce transgenic offspring. Gene manipulation of FGSCs is a rapid and efficient method of animal transgenesis and may serve as a powerful tool for biomedical science and biotechnology.
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