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    • 4. 发明授权
    • Initiation of full mammalian oocyte activation by multiple sperm components
    • 通过多种精子成分启动完全哺乳动物卵母细胞活化
    • US06593139B1
    • 2003-07-15
    • US09478848
    • 2000-01-07
    • Ryuzo YanagimachiAnthony C. F. Perry
    • Ryuzo YanagimachiAnthony C. F. Perry
    • C12N500
    • C12N5/0609C07K14/4705C12N2500/84C12N2517/10
    • The invention provides a microinsertion-based, trans-complementation method of in vitro oocyte activation useful to identify properties of sperm-borne oocyte-activating factor(s) (SOAF) and ooplasmic interactions with sperm components at fertilization. The invention provides at least one detergent-insoluble heat-sensitive component of the perinuclear matrix of a spermatozoon (SOAFm) that acts coordinately with at least one heat-stable submembrane sperm component. SOAFm is solubilized in vitro (SOAFs) under reducing conditions similar to those encountered in the ooplasm. By the method of the invention, the failure of heat-inactivated demembranated sperm heads to activate an egg is rescued by coinsertion of SOAFs into an oocyte. SOAFs is protease-sensitive and is liberated from demembranated heads in a temperature-dependent manner that inversely correlates with the ability of demembranated sperm heads to activate oocytes.
    • 本发明提供了体外卵母细胞激活的基于微插入的反互补方法,其用于鉴定精子传播的卵母细胞活化因子(SOAF)的性质以及在受精时与精子成分的卵泡相互作用。 本发明提供了至少一种与至少一种热稳定的亚膜精子成分协同作用的精子(SOAFm)的核周基质的至少一种不溶于洗涤剂的热敏成分。 SOAFm在与卵泡中遇到的相似的还原条件下在体外溶解(SOAF)。 通过本发明的方法,通过将SOAFs插入卵母细胞而获得热灭活的精子头激活卵的失败。 SOAF是蛋白酶敏感的,并且以温度依赖的方式从分化的头部释放,与分化的精子头激活卵母细胞的能力成反比关系。
    • 8. 发明授权
    • Cumulus cells as nuclear donors
    • 积细胞作为核供体
    • US06331659B1
    • 2001-12-18
    • US09132104
    • 1998-08-10
    • Teruhiko WakayamaRyuzo Yanagimachi
    • Teruhiko WakayamaRyuzo Yanagimachi
    • C12N1500
    • C12N15/8775
    • Animals are produced following injection of adult somatic cell nuclei into enucleated oocytes. The invention provides a method for cloning an animal by directly inserting at least a portion of the adult somatic nucleus (including the minimum chromosomal material able to support development) into a recipient enucleated oocyte. Preferably, the nucleus is inserted by microinjection and, more preferably, by piezo electrically-actuated microinjection. The oocyte is activated prior to, during, or up to about 6 hours after insertion of the nucleus, by electroactivation or exposure to a chemical activating agent, such as Sr2+. The activated renucleated oocyte is allowed to develop into an embryo and is transplanted to a host surrogate mother to develop into a live offspring.
    • 在将成体体细胞核注射到去核卵母细胞后产生动物。 本发明提供了一种用于通过将至少一部分成体体细胞核(包括能够支持发育的最小染色体材料)直接插入受体去核卵母细胞来克隆动物的方法。 优选地,通过显微注射插入细胞核,更优选地,通过压电电致动显微注射。 卵母细胞在插入核后之前,期间或至多约6小时期间被激活,通过电活化或暴露于化学活化剂如Sr2 +。 将激活的复核卵母细胞发育成胚胎,并移植到宿主代孕母体中以形成活的后代。