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    • 3. 发明公开
    • 유선상피세포의 분화 유도용 조성물 및 이를 이용한 유선상피세포의 분화 유도 방법
    • 用于诱导上皮细胞分化的组合物和使用组合物诱导上皮细胞分化的方法
    • KR1020130109857A
    • 2013-10-08
    • KR1020120031957
    • 2012-03-28
    • 충북대학교 산학협력단
    • 김남형이화영허영태이승은엄상준
    • C12N5/071C12N5/02C12N15/63
    • PURPOSE: A composition for inducing differentiation of mammary epithelial cells is provided to establish mammary alveolar cells (MAC-T) in which the expression of casein gene is increased by inducing differentiation of MAC-T and to establish a validation system for identifying the expression and the production of a target gene. CONSTITUTION: A composition for inducing differentiation of mammary epithelial cells contains retinoic acid as an active ingredient. The composition additionally contains prolactin. The mammary epithelial cells are derived from an animal selected among cattle, goat, and sheep. A method for inducing differentiation of mammary epithelial cells comprises the steps of: preparing mammary epithelial cells; and culturing the mammary epithelial cells in a medium containing retinoic acid as an active ingredient.
    • 目的:提供一种用于诱导乳腺上皮细胞分化的组合物,以建立乳腺泡细胞(MAC-T),其中通过诱导MAC-T的分化来增加酪蛋白基因的表达,并建立鉴定表达和 生产目标基因。 构成:用于诱导乳腺上皮细胞分化的组合物含有视黄酸作为活性成分。 该组合物另外含有催乳素。 乳腺上皮细胞衍生自选自牛,山羊和绵羊的动物。 诱导乳腺上皮细胞分化的方法包括以下步骤:制备乳腺上皮细胞; 并在含有视黄酸作为活性成分的培养基中培养乳腺上皮细胞。
    • 7. 发明公开
    • 소 복제 수정란의 체외배양 방법
    • 在体育文化中改善基因组织胚胎
    • KR1020110050100A
    • 2011-05-13
    • KR1020090106944
    • 2009-11-06
    • 충북대학교 산학협력단
    • 김남형추이시앙쑨쉬융난센싱후이
    • C12N5/07C12N5/02
    • PURPOSE: A method for in vitro culture of bovine cloned fertilized eggs is provided to enhance cell generation rate. CONSTITUTION: A method for in vitro culture of bovine cloned fertilized eggs comprises: a step of preparing a culture medium in stock form and storing in a refrigerator or freezer; a step of treating 50nM of trichostatin A to the basic culture medium under the presence of 0.4% BSA(bovine serum albumin); a step of culturing the bovine cloned fertilized eggs in vitro; a step of isolating cells from bovine ear and establishing somatic cell line; a step of removing cumulus cells using hyaluronidase for enucleation; a step of pricking the zona pellucida using a enucleation pipette and inhaling cytoplasm and first polar body; a step of transplanting a donor cell to the enucleated egg; a step of injecting one donor cell; and a step of fusing and activating.
    • 目的:提供牛克隆受精卵的体外培养方法,以提高细胞产生率。 构成:牛克隆受精卵的体外培养方法包括:制备储备形式的培养基并储存在冰箱或冷冻机中的步骤; 在0.4%BSA(牛血清白蛋白)存在下将50nM曲古抑菌素A处理至碱性培养基的步骤; 体外培养牛克隆受精卵的步骤; 从牛耳分离细胞并建立体细胞系的步骤; 使用透明质酸酶去除卵丘细胞进行摘除的步骤; 使用去核移液管穿刺透明带并吸入细胞质和第一极体的步骤; 将供体细胞移植到去核卵的步骤; 注入一个供体细胞的步骤; 和融合和激活的一个步骤。
    • 10. 发明公开
    • 간편하고 효과적인 PAT 분석방법
    • 简单有效的聚合测试方法
    • KR1020150123398A
    • 2015-11-04
    • KR1020140049404
    • 2014-04-24
    • 충북대학교 산학협력단
    • 김남형임자력허영태최향순남궁석엄상준
    • C12Q1/68
    • C12Q1/686C12Q1/6806
    • 본발명은신규한 PAT 분석방법에관한것으로, (a) 서열번호 1의염기서열을갖는앵커프라이머를디자인하는단계; (b) 분석하고자하는목적유전자로부터전사된 RNA에특이적으로결합할수 있는 RNA 특이적프라이머를디자인하는단계;(c) 분석하고자하는목적유전자로부터전사된 RNA를주형으로하고인산화된올리고 dT를첨가하여 65℃에서 10분간반응시킨후, 5x SuperscriptII RT 버퍼용액, 0.1M DTT , 10mM dNTPs, 10mM dATP, 10U/ul T4 DNA 라이게이즈및 DEPC가처리된물을함유한반응액을첨가하여 42℃의온도에서 20분간반응시키는단계; (d) 반응이완료된반응액에서열번호 1의염기서열을갖는앵커프라이머를첨가하여라이게이션반응시킨후, 역전사효소를사용하여 cDNA를합성하는단계; 및 (e) 합성된 cDNA를주형으로하여상기앵커프라이머및 RNA 특이적프라이머를첨가하여 PCR 반응을수행하는단계를포함한다. 본발명에서고안한 PAT(poly A tail)분석방법은오랜시간과복잡한과정에의해수행되던종래 PAT 어세이방법을간소화시켜검출시간및 비용을절감시킬수 있는새로운 PAT 어세이방법을제공하는효과가있다.
    • 本发明涉及一种新的聚A尾(PAT)分析方法,包括以下步骤:(a)设计具有SEQ ID NO:1的碱基序列的锚定引物; (b)设计能够与待分析的靶基因转录的RNA特异性结合的RNA特异性引物; (c)使用从靶基因转录的RNA作为模具进行分析,加入磷酸化的寡聚dT,使其在65℃下反应10分钟,加入包含5x SuperscriptII逆转录酶(RT)缓冲液的反应液, 0.1M DTT,10mM dNTP,10mM dATP,10U /μlT4 DNA连接酶和DEPC处理的水,并使其在42℃下反应20分钟; (d)在反应完成的反应液中加入具有SEQ ID NO:1的碱基序列的锚定引物,进行连接,并使用逆转录酶合成cDNA; 和(e)使用合成的cDNA作为模具,加入锚定引物和RNA特异性引物,并进行PCR。 本发明设计的PAT分析方法简化了通过复杂程序长时间进行的常规PAT测定方法,因此可以提供能够减少检测时间和成本的新型PAT测定方法。