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11.发明授权
公开(公告)号：KR100267668B1
公开(公告)日：2000-11-01
申请号：KR1019980016309
申请日：1998-05-07
IPC: C12N15/54
Abstract: PURPOSE: A serine palmitoyl transferase coding gene derived from Pichia ciferrii and a tetraacetyl phytosphingosine(TAPS) producing method using the same are provided, thereby tetraacetyl phytosphingosine(TAPS) can be mass produced with low costs. CONSTITUTION: The gene LCB2 encoding serine palmitoyl transferase and derived from Pichia ciferrii (ATCC 14091) is represented by sequence ID No. 1 described as in the description. The expression vector prACL2(KCTC-0468BP) having an endonuclease map of Fig. 3 contains the gene LCB2, CYHr gene being resistant to cycloheximide, and ribosome DNA derived from Pichia ciferrii, wherein the ribosome DNA comprises a 0.6kb non transcribed region which is obtained by cleavage with HindIII/EcoRV. A transformant is produced by transforming Pichia ciferrii with the expression vector prACL2(KCTC-0468BP). Tetraacetyl phytosphingosine(TAPS) which is a raw material of ceramides is produced by incubating the transformed Pichia ciferrii and extracting TAPS from the fermented culture, wherein ceramides are used in protecting the skin and maintaining the humidity of the skin.
Abstract translation: 目的：提供从毕赤酵母（Pichia ciferrii）衍生的丝氨酸棕榈酰转移酶编码基因和使用其的四乙酰植物鞘氨醇（TAPS）生产方法，从而可以低成本批量生产四乙酰植物鞘氨醇（TAPS）。 构成：编码丝氨酸棕榈酰转移酶并衍生自毕赤酵母（ATCC 14091）的基因LCB2由描述中描述的序列ID 1表示。 具有图1的核酸内切酶图谱的表达载体prACL2（KCTC-0468BP） 3包含基因LCB2，对放线菌酮具有抗性的CYHr基因和衍生自毕赤酵母的核糖体DNA，其中核糖体DNA包含通过用HindIII / EcoRV切割获得的0.6kb非转录区域。 通过用表达载体prACL2（KCTC-0468BP）转化毕赤酵母来产生转化体。 作为神经酰胺原料的四乙酰植物鞘氨醇（TAPS）是通过将转化的毕赤酵母培养并从发酵培养物中提取TAPS产生的，其中使用神经酰胺来保护皮肤并保持皮肤湿度。
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公开(公告)号：KR100267666B1
公开(公告)日：2000-11-01
申请号：KR1019980016310
申请日：1998-05-07
Abstract: PURPOSE: Provided is an expression plasmid for Pichia ciferrii to insert targeted genes as many as possible into the strain when transformation of Pichia ciferrii is required. CONSTITUTION: The expression plasmid for Pichia ciferrii is obtained by the next steps of: i) separating the gene of ribosome protein L41 of sequence ID. No. 1 from Pichia ciferrii; ii) making the L41 gene resistant to cycloheximide and manufacturing plasmid pCYH1.9¬r to select transformants; iii) separating ribosome DNA and making plasmid prDX9.0 to insert targeted genes into chromosome more effectively; and iv) making a recombinant plasmid by treating ribosome DNA with restriction enzyme.
Abstract translation: 目的：提供毕赤酵母在需要转化毕赤酵母时将目标基因尽可能多地插入菌株的表达质粒。 构成：通过以下步骤获得毕赤酵母的表达质粒：i）分离序列ID的核糖体蛋白L41的基因。 毕赤酵母1号; ii）使L41基因对放线菌酮具有抗性，并制备质粒pCYH1.9-r以选择转化体; iii）分离核糖体DNA，制作质粒prDX9.0，将靶基因更有效地插入染色体; 和iv）用限制酶处理核糖体DNA制备重组质粒。
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公开(公告)号：KR100141651B1
公开(公告)日：1998-06-15
申请号：KR1019940022499
申请日：1994-09-07
CPC classification number: C07K14/815 , Y10S530/855
Abstract: 본 발명은 히루딘을 히루딘을 정제하는 방법을 제공한다. 본 발명의 방법은 조(粗) 히루딘액을 금속 이온으로서 구리이온을 사용하고 용리액으로서 인산염 완충액을 사용하는 금속이온 친화성 크로마토그래피를 이용하여 정제함을 특징으로 한다. 본 발명의 방법에 의하면 종래의 히루딘의 정제방법에 비해 저렴하고 간단하게 히루딘을 고순도로 정제할 수 있는 장점이 있다.
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15.发明授权피치아 시페리 유래의 글리세르알데히드-3-인산 탈수소효소 프로모터 유전자 및 이를 이용한 TAPS 생산방법 失效衍生自Pichiaceperi的甘油醛-3-磷酸脱氢酶启动子基因和使用其制备TAPS的方法
公开(公告)号：KR100287483B1
公开(公告)日：2001-05-02
申请号：KR1019980033969
申请日：1998-08-21
Abstract: 본 발명은
피치아 시페리 (
Pichia ciferrii ) ATCC 14091 유래의 글리세르알데히드-3-인산 탈수소효소(GAPDH : glyceraldehyde-3-phosphate dehydrogenase) 프로모터 유전자, 이 프로모터 유전자를 포함하는 플라스미드 prACGL2(KCTC-0498BP)와 prHECGL2(KCTC-0511BP), 이에 의해 형질전환된
피치아 시페리 형질전환체 및 이를 배양하여 세라미드의 원료인 테트라아세틸 피토스핑고신(TAPS : tetraacetyl phytosphingosine)을 대량 생산하는 방법을 제공한다.
본 발명에 의해 제공되는 플라스미드 prACGL2(KCTC-0498BP) 또는 prHECGL2(KCTC-0511BP)에 의해 형질전환된 형질전환체는 모균주 KFCC-10937에 비하여 최소 2.1배의 TAPS 생산능을 갖고 있다.-
Patent Agency Ranking
公开(公告)号：KR100256023B1
公开(公告)日：2000-05-01
申请号：KR1019970054926
申请日：1997-10-24
Applicant: 한국과학기술연구원
Abstract: PURPOSE: Provided are an expression vector for Hansenula polymorpha which introduces foreign genes in multiple rows and a method for easily selecting Hansenula polymorpha transformants. CONSTITUTION: An expression vector for Hansenula polymorpha is inserted into cells in multiple rows. At 3'-terminal, the vector has repeated segments of 5'-GGGTGGCG-3', self-replicable segment, promoter, and selective genes for nutritional requirement etc. And a method for selecting Hansenula polymorpha transformants is comprised of the next steps of: i) transforming Hansenula polymorpha with the expression vector; ii) cultivating and selecting transformants in a minimal medium; iii) cultivating the transformants continuously in a complex medium to stabilize vectors; and iv) cultivating stabilized transformants in a selective medium containing antibiotics to select transformants resistant to the antibiotics.
Abstract translation: 目的：提供多种多形汉逊酵母的表达载体，其可以将多个外来基因引入多行，并且可以方便地选择汉逊酵母多形汉逊酵母转化株。 构成：将多形汉逊酵母的表达载体插入多行细胞。 在3'末端，载体具有重复的5'-GGGTGGCG-3'区段，可自我复制区段，启动子和选择性基因的营养需求等。选择汉逊酵母多形汉逊酵母转化体的方法包括以下步骤 ：i）用表达载体转化汉逊酵母多形汉逊酵母; ii）在基本培养基中培养和选择转化体; iii）在复杂培养基中连续培养转化体以稳定载体; 和iv）在含有抗生素的选择性培养基中培养稳定的转化体以选择对抗生素具有抗性的转化体。
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公开(公告)号：KR100252787B1
公开(公告)日：2000-04-15
申请号：KR1019970054925
申请日：1997-10-24
Applicant: 한국과학기술연구원
Abstract: PURPOSE: Provided are multiple tandem integrating autonomously replicating sequence(ARS) originated from Hansenula polymorpha and an expression vector including the same. The vector including this sequence is very useful for the production of an exogenous protein from Hansenula polymorpha due to its high efficiency and stability of transformation. CONSTITUTION: A vector for inserting polynucleotide coding hetero protein into chromosome of Hansenula polymorpha is composed of the followings: (a) multiple tandem integrating ARS including repetitive sequence of 5'-GGGTGGCG-3' at 3' terminal originated from Hansenula polymorpha DL-1(ATCC 26012); (b) a promotor; (c) polynucleotide coding hetero protein located at downstream of the promotor; and (d) a terminator located at downstream of the nucleotide.
Abstract translation: 目的：提供源自多形汉逊酵母的多重串联整合自主复制序列（ARS）和包含其的表达载体。 包含该序列的载体由于其转化的高效率和稳定性而非常适用于从多形汉逊酵母（Hansenula polymorpha）生产外源蛋白质。 构成：用于将多核苷酸编码杂蛋白插入多形汉逊酵母染色体的载体由以下组成：（a）多重串联整合ARS，包括来自多形汉逊酵母DL-1的3'末端的5'-GGGTGGCG-3'的重复序列 （ATCC 26012）; （b）促进者; （c）位于启动子下游的编码杂蛋白的多核苷酸; 和（d）位于核苷酸下游的终止子。
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18.发明公开피치아 시페리 유래의 글리세르알데히드-3-인산 탈수소효소 프로모터 유전자 및 이를 이용한 TAPS 생산방법 失效从PICHIA CIFERRII衍生的甘油三磷酸脱氢酶促进剂基因及其制备方法
公开(公告)号：KR1020000014502A
公开(公告)日：2000-03-15
申请号：KR1019980033969
申请日：1998-08-21
Abstract: PURPOSE: Glyceraldehyde-3-phsphate dehydrogenase (GAPDH) promoter gene isolated from GAPDH gene coding glyceraldehyde-3-phsphate dehydrogenase derived from pichia ciferrii is able to increase yield of tetraacetyl phytosphingosine(TAPS) by amplifying LCB2 gene. CONSTITUTION: Tetraacetyl phytosphingosine(TAPS) is produced by cultivating transformant pichia ciferrii transformed with plasmid containing ribosome DNA derived from pichia ciferrii, CYH gene with resistance to antibiotic cycloheximide, glyceraldehyde-3-phsphate dehydrogenase(GAPDH) promoter gene, LCB2 gene coding serine palmitoyl transferase derived from pichia ciferrii and by extracting tetraacetyl phytosphingosine from culture media.
Abstract translation: 目的：从GAPDH基因中分离得到的甘油醛-3-磷酸脱氢酶（GAPDH）启动子基因，通过扩增LCB2基因能够提高四氢乙酰植物鞘氨醇（TAPS）的产量。 构成：四乙酰植物鞘氨醇（TAPS）是通过培养由含有来自毕赤酵母的核糖体DNA，具有抗生素环己酰亚胺，甘油醛-3-磷酸脱氢酶（GAPDH）启动子基因，编码丝氨酸棕榈酰基的LCB2基因的CYH基因的质粒转化的转化体毕赤酵母产生的 从毕赤酵母衍生的转移酶和从培养基中提取四乙酰植物鞘氨醇。
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19.发明公开인간 혈장 알부민 발현벡터를 함유한 형질전환 효모 사카로마이세스 세레비시에를 이용한 인간 혈장 알부민의 제조 방법 失效通过使用含有人血清白蛋白转移载体的转化YEAST SACCHAROMYCES认证制备人血清白蛋白
公开(公告)号：KR1020000010068A
公开(公告)日：2000-02-15
申请号：KR1019980030775
申请日：1998-07-30
Applicant: 한국과학기술연구원 , 제이더블유중외제약 주식회사
IPC: A61K38/38
CPC classification number: C07K14/765 , C12N1/16 , C12N15/81
Abstract: PURPOSE: A method is provided to culture and generate efficiently a human serum albumin by minimizing the analysis of the albumin being revealed from a transformed body. CONSTITUTION: A method for generating a re-combined human serum albumin comprises: assembling a revealing vector containing an existent promoter being connected to a human serum albumin cDNA to a yeast saccharomyces cerevisiae; introducing the assembled material to the yeast saccharomyces cerevisiae for forming a transformed yeast strain; fermenting and culturing for revealing efficiently the human serum albumin. Herein, a buffering liquid or a nitrogen is added to the transformed yeast strain for maintaining the pH of the culturing for 5 ¯ 6 for minimizing the analysis of the albumin being revealed by fermenting and culturing.
Abstract translation: 目的：提供一种通过最小化从转化体发现的白蛋白的分析来培养和有效生成人血清白蛋白的方法。 构成：产生重组合人血清白蛋白的方法包括：将含有与人血清白蛋白cDNA连接的存在启动子的显示载体装配到酵母酿酒酵母; 将组装的材料引入到酵母酿酒酵母中以形成转化的酵母菌株; 发酵培养，有效显露人血清白蛋白。 本文中，向转化的酵母菌株中加入缓冲液或氮气以保持培养物的pH值为5，以最小化通过发酵和培养所揭示的白蛋白的分析。
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公开(公告)号：KR100237979B1
公开(公告)日：2000-01-15
申请号：KR1019970054927
申请日：1997-10-24
Applicant: 한국과학기술연구원
IPC: C12N15/81
Abstract: 본 발명은 한세눌라 폴리모르파(Hansenula polymorpha)의 글리세르알데히드-3-인산 탈수소효소(GAPDH) 유전자, GAPDH 프로모터 및 이 프로모터를 함유하는 발현 벡터에 관한 것으로, 본 발명에 따른 GAPDH 프로모터는 강력한 구성적 발현 프로모터로서 탄소원에 따른 별도의 발현 유도가 필요하지 않으므로 한세눌라 폴리모르파에서 외래 단백질을 발현시키는데 매우 유용하다.
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