公开(公告)号：KR1020120067534A

公开(公告)日：2012-06-26

申请号：KR1020100128984

申请日：2010-12-16

Applicant: 한국화학연구원

Inventor： 이우길 ,  이종교 ,  김미현 ,  김해수

IPC: C12N15/10 ,  C12N15/51 ,  C12Q1/68 ,  C12N9/14

CPC classification number: C12N15/1003 ,  C12N9/14 ,  C12Q1/6806 ,  C12Q1/706 ,  G01N33/56983

Abstract: PURPOSE: A method for isolating genome DNA from hepatitis B virus(HBV) is provided to screen a therapeutic agent for HBV and to enable genome DNA quantitation. CONSTITUTION: A method for isolating genome DNA from HBV comprises: a step of adding cell lysis material to cells which produces HBV and centrifuging to prepare supernatant; a step of adding protease to the supernatant; and a step of adding phenol/chloroform/isoamyl alcohol to a lysate to prepare genome DNA. The cells are HepG2.2.15, HepW10 or HepD2. The cell lysis material is a RIPA solution containing 50 mM Tris-HCl(pH 8.0), 0.5 M EDTA, 150 mM NaCl, 1% NP-40 and 1 mM phenylmethansulfonyl fluoride(PMSF). The protease is proteinase K. A method for quantitation of the genome DNA includes PCR, southern blotting, or dot blotting.

Abstract translation: 目的：提供一种从乙型肝炎病毒（HBV）分离基因组DNA的方法，以筛选HBV的治疗剂并使基因组DNA定量。 构成：从HBV分离基因组DNA的方法包括：向产生HBV的细胞中加入细胞裂解物质并离心制备上清液的步骤; 向上清液中加入蛋白酶的步骤; 以及向裂解物中加入苯酚/氯仿/异戊醇以制备基因组DNA的步骤。 细胞为HepG2.2.15，HepW10或HepD2。 细胞裂解物质是含有50mM Tris-HCl（pH8.0），0.5M EDTA，150mM NaCl，1％NP-40和1mM苯基甲磺酰氟（PMSF）的RIPA溶液。 蛋白酶是蛋白酶K.用于定量基因组DNA的方法包括PCR，Southern印迹或斑点印迹。

公开(公告)号：KR1020130116155A

公开(公告)日：2013-10-23

申请号：KR1020120118708

申请日：2012-10-24

Applicant: 주식회사 강스템바이오텍 ,  서울대학교산학협력단

Inventor： 강경선 ,  서광원 ,  김미현

IPC: C12N5/071 ,  C12N15/85 ,  C12N5/10 ,  A61K35/12

Abstract: PURPOSE: A method for directly producing functional insulin-producing cells from skin cells is provided to directly convert human somatic cells into insulin-producing cells without dedifferentiation into pluripotent cells and to obtain the insulin-producing cells which normally secrete insulin by glucose stimulation. CONSTITUTION: A method for producing insulin-producing cells comprises the step of promoting transcription of PDX-1 gene in somatic cells which do not produce insulin. The method directly converts the somatic cells into insulin-producing cells without dedifferentiation into cells with stemness. The step for promoting transcription of PDX-1 gene comprises the steps of transfecting cells by SMAD2 and transducing anti-Let7i by culturing the transfected cells for 2-10 days. [Reference numerals] (AA) Smad2 trait infection; (BB) Second day; (CC) FGM medium + 1% BSA; (DD) Anti-Let7i trait infection; (EE) EGM medium + 1% BSA; (FF) Seventh day; (GG) Glucose simulation index

Abstract translation: 目的：提供从皮肤细胞直接产生功能性胰岛素生成细胞的方法，以将人体细胞直接转化成胰岛素产生细胞，而不去分化成多能细胞，并获得通过葡萄糖刺激通常分泌胰岛素的胰岛素产生细胞。 构成：产生胰岛素的细胞的方法包括促进PDX-1基因在不产生胰岛素的体细胞中的转录的步骤。 该方法直接将体细胞转化为产生胰岛素的细胞，而无需去分化成具有干细胞的细胞。 促进PDX-1基因转录的步骤包括通过SMAD2转染细胞并通过培养转染细胞2-10天来转导抗Let7i的步骤。 （附图标记）（AA）Smad2性状感染; （BB）第二天; （CC）FGM培养基+ 1％BSA; （DD）抗let7i性状感染; （EE）EGM培养基+ 1％BSA; （FF）第七天; （GG）葡萄糖模拟指数