-
公开(公告)号：KR100358754B1
公开(公告)日：2002-11-07
申请号：KR1020000006888
申请日：2000-02-14
Applicant: 대한민국(농촌진흥청장)
IPC: C12N15/00
Abstract: 본 발명은 유전공학적인 방법인 유전자 재조합 기술을 이용하여 돼지의 유선에서 인간의 조혈촉진제(EPO:erythropoietin)를 대량으로 생산할 수 있는 형질전환 돼지
(TRANSGENIC PORCINE) 를 생산하는 것에 관한 것으로서, 더 상세하게는 인간의 신장에 존재하는 조혈촉진제(EPO) 유전자를 돼지의 수정란에 주입, 형질전환시키므로서 인간의 조혈촉진제(EPO)유전자를 가지는 돼지(새롬이)를 생산하고 그 돼지의 젖을 통하여 고가의 의약품인 인간의 조혈촉진제를 대량으로 분비할 수 있도록 하므로서 인류건강의 진일보에 기여함은 물론 고가의 의약품 수출을 통하여 국가경제 및 국가경쟁력 기반구축에 크게 기여할 수 있는 특성을 가지는 발명이다.
이에 본 발명의 인간의 조혈촉진제(EPO) 생산을 위한 형질전환 돼지의 생산은 유전자 염기서열이 도 3와 같이 구성되는 것을 특징으로 한다.-
公开(公告)号：KR1020010018289A
公开(公告)日：2001-03-05
申请号：KR1019990034180
申请日：1999-08-18
Applicant: 대한민국(농촌진흥청장) , 주식회사 녹십자홀딩스
IPC: G01N33/48
Abstract: PURPOSE: A pregnancy diagnosis kit is provided to readily decide the pregnancy of a cow 19 or 20 days after fertilization by using the concentration of progesterone existing in blood. CONSTITUTION: A thin film type paper(1) forms a specific size of pores inside. A progesterone-BSA conjugate(2) of specific concentration is coated in a specific area on at least one face of the paper. A colloidal gold label(3) embedding anti-progesterone is combined with some part of an upper face of the progesterone-BSA conjugate. The concentration of progesterone existing in blood, progesterone-cow serum albumin fixed on a thin film and an anti-progesterone gold conjugate are competitively reacted. Color reaction is performed in reverse proportion to the progesterone concentration in the blood. Accordingly, the pregnancy of the cow is readily decided.
Abstract translation: 目的：使用怀孕诊断试剂盒，通过使用血液中存在的孕酮浓度，容易地在受精后19或20天确定奶牛的怀孕。 构成：薄膜型纸（1）内部形成特定尺寸的孔。 将特定浓度的孕酮-BSA结合物（2）涂布在纸的至少一个面上的特定区域中。 胶体金标记（3）嵌入抗孕酮与孕酮-BSA结合物的上表面的一部分组合。 存在于血液中的孕酮的浓度，固定在薄膜上的孕酮 - 牛血清白蛋白和抗孕酮金缀合物的竞争性反应。 彩色反应与血液中的孕酮浓度成反比。 因此，牛的怀孕很容易决定。
-
公开(公告)号：KR100375672B1
公开(公告)日：2003-03-15
申请号：KR1019990032079
申请日：1999-08-05
Applicant: 대한민국(농촌진흥청장)
IPC: C07K14/59
Abstract: PURPOSE: Provided is a method for manufacturing a single chain bovine Follicle-stimulating hormone(FSH) which shows a stronger hormone activity than one of natural hormones, so as to be used for animal drugs. Thereby, FSH is easily manufactured in a high yield as forming FSH consisting of alpha and beta short fragments, into single chain FSH. CONSTITUTION: A method for manufacturing a single chain bovine Follicle-stimulating hormone(FSH) is characterized by the following steps of: i) performing polymerase chain reaction to link DNA coding an alpha short fraction of bovine FSH to DNA coding a beta short fragment of the hormone, using 4kinds of oligonucleic acids bases as primers, to obtain cDNAs of the alpha and beta fractions and cloning the cDNAs with pUC119; ii) multiplying cloned DNA through PCR to obtain bFSH composed of cDNA of alpha and beta fractions, cutting the bFSH by restriction enzyme Kpnl/Xbal and linking the bFSH to vector pUC119 to form expression vector pcDAN3-bFSH; iii) inserting the expression vector pcDAN3-bFSH into CHO-K1 and cultivating CHO-K1 in a medium Ham F-12 including penicillin, streptomycin, glutamine, 10FCS and G418, in the presence of 5CO2 and air, at 37 deg.C for 2 weeks and selecting stable cells: iv) cultivating selected cells at 20 ml of CHO-S-SFM-11 medium containing 50 units/ml of penicillin and 50 microg/ml of streptomycin at 37 deg.C for 48 hours, and centrifuging supernatant liquid obtained through the cultivation for 60 minutes to remove cell remnants.
Abstract translation: 目的：提供一种生产单一链牛卵泡刺激素（FSH）的方法，其表现出比天然激素更强的激素活性，以用于动物药物。 因此，FSH很容易以高产率生产，形成由α和β短片段组成的FSH，进入单链FSH。 构成：制造单链牛卵泡刺激素（FSH）的方法的特征在于以下步骤：i）进行聚合酶链式反应以将编码牛FSH的α短部分的DNA与编码β短片段的DNA 使用4种寡核苷酸碱基作为引物，获得α和β部分的cDNA并用pUC119克隆cDNA; ii）通过PCR扩增克隆的DNA以获得由α和β部分的cDNA组成的bFSH，通过限制性内切酶KpnI / XbaI切割bFSH并将bFSH与载体pUC119连接以形成表达载体pcDAN3-bFSH; iii）将表达载体pcDAN3-bFSH插入到CHO-K1中，并且在37℃下在含有青霉素，链霉素，谷氨酰胺，10FCS和G418的培养基Ham F-12中，在5CO2和空气存在下培养CHO-K1 2周并选择稳定的细胞：iv）在20ml包含50单位/ ml青霉素和50μg/ ml链霉素的CHO-S-SFM-11培养基中于37℃培养选择的细胞48小时，并离心上清液 通过培养60分钟获得的液体除去细胞残留物。
-
公开(公告)号：KR1020010081456A
公开(公告)日：2001-08-29
申请号：KR1020000006888
申请日：2000-02-14
Applicant: 대한민국(농촌진흥청장)
IPC: C12N15/00
CPC classification number: A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/108 , A01K2267/01 , C12N15/8509
Abstract: PURPOSE: A method for producing transgenic porcines for producing human erythropoietin(EPO) is provided, thereby human erythropoietin(EPO) can be mass-produced. CONSTITUTION: The method for producing transgenic porcines comprising the steps of: cloning the genomic DNA of human erythropoietin(EPO) using a WAP promoter isolated from the mammary gland of mouse by polymerase chain reaction(PCR); preparing the EPO expression vector containing human erythropoietin(hEPO) and SV40 poly A gene; injecting eCG hormone and hCG hormone into the fertilized eggs of porcine to induce overovulation; recovering the fertilized eggs and inserting the above DNA into a male cell nucleus using a manipulator; transplanting the cell into a surrogate porcine mother; and producing transgenic porcines having DNA represented by SEQ ID NO: 1 from the surrogate porcine mother.
Abstract translation: 目的：提供生产人促红细胞生成素（EPO）的转基因猪的方法，从而大量生产人促红细胞生成素（EPO）。 构成：生产转基因猪的方法，包括以下步骤：使用通过聚合酶链反应（PCR）从小鼠乳腺分离的WAP启动子克隆人促红细胞生成素（EPO）的基因组DNA; 制备含有促红细胞生成素（hEPO）和SV40聚A基因的EPO表达载体; 将eCG激素和hCG激素注射到猪的受精卵中以诱导过度排卵; 回收受精卵，并使用机械手将上述DNA插入雄性细胞核; 将细胞移植到替代猪母亲中; 并从代孕猪母体生产具有由SEQ ID NO：1表示的DNA的转基因猪。
-
公开(公告)号：KR100375671B1
公开(公告)日：2003-03-15
申请号：KR1019990032078
申请日：1999-08-05
Applicant: 대한민국(농촌진흥청장)
IPC: C07K14/59
Abstract: PURPOSE: Provided is a method for manufacturing single chain bovine luteinizing hormone(LH) which shows stronger hormone activity than one of natural LH, so as to be used for animal drugs. Thereby, LH is easily manufactured in a high yield as forming LH consisting of alpha and beta short fragments, into single chain LH. CONSTITUTION: A method for manufacturing single chain bovine luteinizing hormone(LH) comprises the following steps of: i) performing polymerase chain reaction to link DNA coding an alpha short fragment of bovine luteinizing hormone(LH) with DNA coding a beta short fragment of the hormone, using 4 kinds of oligonucleic acids bases as primers, to obtain cDNAs of the alpha and beta short fragments and cloning the cDNAs with pUC119; ii) multiplying cloned DNA through PCR to obtain bLH composed of cDNA of alpha and beta fragments, cutting the bLH by restriction enzyme Kpnl/Xbal and linking the bLH to vector pUC119 to form expression vector pcDAN3-bLH; iii) inserting the expression vector pcDAN3-bLH into CHO-K1 and cultivating CHO-K1 at media Ham F-12 including penicillin, streptomycin, glutamine, 10FCS and G418, in the presence of 5CO2 and air, at 37 deg.C for 2 weeks and selecting stable cells: iv) cultivating selected cells at 20 ml of CHO-S-SFM-11 media containing 50 units/ml of penicillin and 50 microg/ml of streptomycin at 37 deg.C for 48 hours, and centrifuging supernatant liquid obtained through the cultivation for 60 minutes to remove cell remnants.
Abstract translation: 目的：提供一种生产单一链牛黄素化激素（LH）的方法，其表现出比天然LH更强的激素活性，以用于动物药物。 由此，容易以高收率制造LH，从而将由α和β短片段组成的LH形成为单链LH。 构成：制造单链牛黄体生成激素（LH）的方法包括以下步骤：i）进行聚合酶链式反应以将编码牛黄体生成素（LH）的α短片段的DNA与编码β链短片段的DNA 激素，以4种寡核苷酸碱基为引物，获得α和β短片段的cDNA，并用pUC119克隆cDNA; ii）通过PCR扩增克隆的DNA以获得由α和β片段的cDNA组成的bLH，通过限制性酶KpnI / XbaI切割bLH并将bLH连接至载体pUC119以形成表达载体pcDAN3-bLH; iii）将表达载体pcDAN3-bLH插入到CHO-K1中，并在37℃下于37℃，在5CO2和空气存在下，在包含青霉素，链霉素，谷氨酰胺，10FCS和G418的培养基Ham F-12中培养CHO-K1 周并选择稳定的细胞：iv）在20ml包含50单位/ ml青霉素和50μg/ ml链霉素的CHO-S-SFM-11培养基中于37℃培养选择的细胞48小时，并离心上清液 通过培养获得60分钟以除去细胞残余物。
-
Patent Agency Ranking
公开(公告)号：KR1020010077538A
公开(公告)日：2001-08-20
申请号：KR1020000005390
申请日：2000-02-03
Applicant: 대한민국(농촌진흥청장)
IPC: A61K38/24
Abstract: PURPOSE: A human chorionic gonadotropin (hcg) consisting of single chain by genetic recombination is provided, which has same physiological activity as natural type human chorionic gonadotropin (hcg), and is thus used for animal medicines and clinical medicines. Also, nucleic acid molecule coding the hormone and its preparation method are provided. CONSTITUTION: A single chain-human chorionic gonadotropin (hcg) is composed of α-unit and β-unit and is identical with or similar to 1-257 site or 21-257 site of sequence list 6. A single chain-human chorionic gonadotropin (hcg) is prepared by the following steps of: (a) amplifying nucleic acids coding α and β units of human chorionic gonadotropin (hgc) by PCR; (b) cloning amplified nucleic acids coding α and β units; (c) coupling the resultant nucleic acids; (d) constructing an expression vector including the coupled nucleic acid; (e) transforming an eukaryotic cell using the expression vector; (f) expressing human chorionic gonadotropin in the eukaryotic cell; and (g) separating the prepared human chorionic gonadotropin.
Abstract translation: 目的：提供由基因重组单链组成的人绒毛膜促性腺激素（hcg），与天然型人绒毛膜促性腺激素（hcg）具有相同的生理活性，因此用于动物药物和临床药物。 另外，提供了编码激素的核酸分子及其制备方法。 构成：单链 - 人绒毛膜促性腺激素（hcg）由α单位和β单元组成，与序列表6的1-257位点或21-257位点相同或相似。单链 - 人绒毛膜促性腺激素 （hcg）通过以下步骤制备：（a）通过PCR扩增编码人绒毛膜促性腺激素（hgc）的α和β单位的核酸; （b）克隆编码α和β单位的扩增核酸; （c）偶联所得核酸; （d）构建包含偶联核酸的表达载体; （e）使用表达载体转化真核细胞; （f）在真核细胞中表达人绒毛膜促性腺激素; 和（g）分离制备的人绒毛膜促性腺激素。
-
公开(公告)号：KR1020010016878A
公开(公告)日：2001-03-05
申请号：KR1019990032078
申请日：1999-08-05
Applicant: 대한민국(농촌진흥청장)
IPC: C07K14/59
Abstract: PURPOSE: Provided is a method for manufacturing single chain bovine luteinizing hormone(LH) which shows stronger hormone activity than one of natural LH, so as to be used for animal drugs. Thereby, LH is easily manufactured in a high yield as forming LH consisting of alpha and beta short fragments, into single chain LH. CONSTITUTION: A method for manufacturing single chain bovine luteinizing hormone(LH) comprises the following steps of: i) performing polymerase chain reaction to link DNA coding an alpha short fragment of bovine luteinizing hormone(LH) with DNA coding a beta short fragment of the hormone, using 4 kinds of oligonucleic acids bases as primers, to obtain cDNAs of the alpha and beta short fragments and cloning the cDNAs with pUC119; ii) multiplying cloned DNA through PCR to obtain bLH composed of cDNA of alpha and beta fragments, cutting the bLH by restriction enzyme Kpnl/Xbal and linking the bLH to vector pUC119 to form expression vector pcDAN3-bLH; iii) inserting the expression vector pcDAN3-bLH into CHO-K1 and cultivating CHO-K1 at media Ham F-12 including penicillin, streptomycin, glutamine, 10FCS and G418, in the presence of 5CO2 and air, at 37 deg.C for 2 weeks and selecting stable cells: iv) cultivating selected cells at 20 ml of CHO-S-SFM-11 media containing 50 units/ml of penicillin and 50 microg/ml of streptomycin at 37 deg.C for 48 hours, and centrifuging supernatant liquid obtained through the cultivation for 60 minutes to remove cell remnants.
Abstract translation: 目的：提供单链牛黄体生成素（LH）的制备方法，其具有比天然LH更强的激素活性，以用于动物药物。 因此，LH容易以高产率制造，形成由α和β短片段组成的LH，形成单链LH。 构成：制备单链牛黄体生成激素（LH）的方法包括以下步骤：i）进行聚合酶链式反应，将编码牛黄体生成激素（LH）的α短片段的DNA与编码β 激素，使用4种寡核苷酸碱基作为引物，获得α和β短片段的cDNA，并用pUC119克隆cDNA; ii）通过PCR扩增克隆的DNA，以获得由α和β片段的cDNA组成的bLH，通过限制酶KpnI / XbaI切割bLH并将bLH与载体pUC119连接以形成表达载体pcDAN3-bLH; iii）将表达载体pcDAN3-bLH插入到CHO-K1中，并在5CO2和空气存在下，在37℃下，在包含青霉素，链霉素，谷氨酰胺，10FCS和G418的培养基Ham-F-12处培养CHO-K1 2 周期并选择稳定细胞：iv）在37℃下，在含有50单位/ ml青霉素和50μg/ ml链霉素的20ml的CHO-S-SFM-11培养基中培养选定的细胞48小时，并将上清液 通过培养获得60分钟去除细胞残留物。
-
公开(公告)号：KR100453571B1
公开(公告)日：2004-10-20
申请号：KR1020010044610
申请日：2001-07-24
Applicant: 대한민국(농촌진흥청장)
IPC: C12N15/85
Abstract: PURPOSE: Provided are a transgenic pig capable of producing human erythropoietin and a preparation method thereof, thereby stably and economically producing a large amount of human erythropoietin from the transgenic pig's urine. CONSTITUTION: A recombinant vector of human erythropoietin, pCR2.1 hEPO, includes EPO gene between KpnI of rat U II promotor having EcoRI and KpnI sites and SalI of SV40 Poly A having SalI and EcoRV sites. The transgenic pig is transformed by the recombinant vector, pCR2.1 hEPO and produces human erythropoietin in its urine.
Abstract translation: 发明目的：提供一种能够产生人促红细胞生成素的转基因猪及其制备方法，由此从转基因猪尿中稳定而经济地生产大量人促红细胞生成素。 构成：人促红细胞生成素的重组载体pCR2.1hEPO包含具有EcoRI和KpnI位点的大鼠UII启动子的KpnI与具有SalI和EcoRV位点的SV40多聚A的SalI之间的EPO基因。 通过重组载体pCR2.1hEPO转化转基因猪并在其尿中产生人促红细胞生成素。
-
公开(公告)号：KR1020010016879A
公开(公告)日：2001-03-05
申请号：KR1019990032079
申请日：1999-08-05
Applicant: 대한민국(농촌진흥청장)
IPC: C07K14/59
Abstract: PURPOSE: Provided is a method for manufacturing a single chain bovine Follicle-stimulating hormone(FSH) which shows a stronger hormone activity than one of natural hormones, so as to be used for animal drugs. Thereby, FSH is easily manufactured in a high yield as forming FSH consisting of alpha and beta short fragments, into single chain FSH. CONSTITUTION: A method for manufacturing a single chain bovine Follicle-stimulating hormone(FSH) is characterized by the following steps of: i) performing polymerase chain reaction to link DNA coding an alpha short fraction of bovine FSH to DNA coding a beta short fragment of the hormone, using 4kinds of oligonucleic acids bases as primers, to obtain cDNAs of the alpha and beta fractions and cloning the cDNAs with pUC119; ii) multiplying cloned DNA through PCR to obtain bFSH composed of cDNA of alpha and beta fractions, cutting the bFSH by restriction enzyme Kpnl/Xbal and linking the bFSH to vector pUC119 to form expression vector pcDAN3-bFSH; iii) inserting the expression vector pcDAN3-bFSH into CHO-K1 and cultivating CHO-K1 in a medium Ham F-12 including penicillin, streptomycin, glutamine, 10FCS and G418, in the presence of 5CO2 and air, at 37 deg.C for 2 weeks and selecting stable cells: iv) cultivating selected cells at 20 ml of CHO-S-SFM-11 medium containing 50 units/ml of penicillin and 50 microg/ml of streptomycin at 37 deg.C for 48 hours, and centrifuging supernatant liquid obtained through the cultivation for 60 minutes to remove cell remnants.
Abstract translation: 目的：提供单链牛卵泡刺激素（FSH）的制备方法，其显示比天然激素更强的激素活性，以用于动物药物。 因此，FSH容易以高产率制造，形成由α和β短片段组成的FSH，成为单链FSH。 构成：用于制造单链牛卵泡刺激素（FSH）的方法的特征在于以下步骤：i）进行聚合酶链反应以将编码牛FSH的α短部分的DNA链接到编码β短链片段的DNA 使用4种寡核苷酸碱基作为引物，获得α和β级分的cDNA，并用pUC119克隆cDNA; ii）通过PCR扩增克隆的DNA，得到由α和β部分的cDNA组成的bFSH，通过限制酶KpnI / XbaI切割bFSH并将bFSH与载体pUC119连接以形成表达载体pcDAN3-bFSH; iii）将表达载体pcDAN3-bFSH插入CHO-K1中，并在5CO2和空气存在下，在37℃下，在包含青霉素，链霉素，谷氨酰胺，10FCS和G418的培养基Ham F-12中培养CHO-K1， 2周，选择稳定细胞：iv）在37℃下，在含有50单位/ ml青霉素和50μg/ ml链霉素的20ml CHO-S-SFM-11培养基中培养选定的细胞48小时，并离心上清液 通过培养60分钟获得的液体去除细胞残留物。
-
公开(公告)号：KR1020030009936A
公开(公告)日：2003-02-05
申请号：KR1020010044610
申请日：2001-07-24
Applicant: 대한민국(농촌진흥청장)
IPC: C12N15/85
CPC classification number: C12N15/8509 , A01K67/0275 , A01K2227/108 , A01K2267/01 , C12N2015/8518
Abstract: PURPOSE: Provided are a transgenic pig capable of producing human erythropoietin and a preparation method thereof, thereby stably and economically producing a large amount of human erythropoietin from the transgenic pig's urine. CONSTITUTION: A recombinant vector of human erythropoietin, pCR2.1 hEPO, includes EPO gene between KpnI of rat U II promotor having EcoRI and KpnI sites and SalI of SV40 Poly A having SalI and EcoRV sites. The transgenic pig is transformed by the recombinant vector, pCR2.1 hEPO and produces human erythropoietin in its urine.
Abstract translation: 目的：提供能够产生人促红细胞生成素的转基因猪及其制备方法，从而稳定且经济地从转基因猪尿中产生大量人促红细胞生成素。 构成：人促红细胞生成素pCR2.1hEPO的重组载体包括具有EcoRI和KpnI位点的大鼠U II启动子的KpnI与具有SalI和EcoRV位点的SV40聚A的SalI之间的EPO基因。 通过重组载体pCR2.1hEPO转化转基因猪，并在其尿液中产生人促红细胞生成素。
-
-
-
-
-
-
-
-
-
Search Results
10
records
(took 0.018 seconds)
