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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。
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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。
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公开(公告)号：KR101911515B1
公开(公告)日：2018-10-25
申请号：KR1020130083027
申请日：2013-07-15
Applicant: 한국생명공학연구원 , 대한민국(농촌진흥청장)
IPC: C12N15/63 , C12N15/85 , A01K67/027
Abstract: 본발명은이종장기이식시에발생할수 있는단계별면역거부반응을억제하는유전자들이알파-1,3-갈락토실트랜스퍼라제(α-1,3-galactosyltransferase) 유전자에적중된복합형질전환벡터가형질전환된복합형질전환세포주및 이의제조방법에관한것으로, 이종이식시 발생하는일련의면역거부반응을한번에조절가능한다중유전자형질전환된세포주를확인함으로써, 다양한인공장기효율적인생산및 복합형질전환복제돼지생산에유용하게이용할수 있다.
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公开(公告)号：KR101514415B1
公开(公告)日：2015-04-27
申请号：KR1020130097300
申请日：2013-08-16
Applicant: 대한민국(농촌진흥청장)
IPC: C07K16/18 , C12N15/13 , A61K39/395 , A61P31/12
Abstract: 본 발명은 닭에서의 면역원성이 감소된 재조합 3D8 scFv(single chain variable fragment) 항체 및 이의 항원 결합부에 관한 것이다. 또한, 본 발명은 상기의 항체 및 이의 항원 결합부를 암호화하는 핵산 분자와 상기의 항체 및/또는 이의 항원 결합부를 포함하는 조성물에 관한 것이다. 본 발명에 따르면 닭에 쥐 유래 3D8 scFv 항체를 주입할 경우 발생할 수 있는 HAMA 유사 현상 등을 유발하지 않도록 면역원성이 감소되며, 핵산결합능, 핵산분해능 및 세포 침투능 등과 같은 기존 3D8 scFv 항체의 활성을 유지하는 재조합 항체가 제공된다.
Abstract translation: 本发明涉及在鸡中免疫原性降低的重组3D8 scFv（单链可变片段）抗体及其抗原结合部分。 本发明还涉及包含编码所述抗体及其抗原结合部分和所述抗体和/或其抗原结合部分的核酸分子的组合物。 根据本发明的免疫原性降低，以便不引起这样的HAMA类似发生，如果鼠3D8 scFv抗体的注射到鸡，保持scFv抗体，现有3D8活性如核酸结合活性，核酸分辨率和细胞chimtuneung Lt。
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公开(公告)号：KR101437590B1
公开(公告)日：2014-09-11
申请号：KR1020120006298
申请日：2012-01-19
Applicant: 대한민국(농촌진흥청장)
IPC: C12N15/63 , C12N15/12 , C12N15/85 , A01K67/027
Abstract: 본 발명은 사람 CD73 발현 벡터, 이를 포함하는 세포주 및 형질전환 복제동물에 관한 것이다. 본 발명에 따르면, 새로운 면역거부반응 조절기술의 개발로 고부가가치의 장기이식용 미니돼지의 생산에 기여할 수 있으며, 새로운 장기 이식용 형질전환 복제 돼지의 생산으로 이종 장기 이식의 실용화를 촉진할 수 있다.
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Patent Agency Ranking
公开(公告)号：KR101359547B1
公开(公告)日：2014-02-12
申请号：KR1020110115226
申请日：2011-11-07
Applicant: 대한민국(농촌진흥청장)
IPC: C12N15/873 , C12N5/10
Abstract: 본 발명은 복제동물의 생산성 향상을 위한 동결융해 공여세포의 처리 방법에 관한 것이다. 보다 상세하게는 급속냉동된 체세포를 융해하여 바로 핵이식에 사용하는 단계를 포함하는 동결융해 공여세포의 처리 방법에 관한 것이다. 본 발명에 따르면 존의 복제동물 생산에서 가장 중요한 과정인 공여세포의 처리 과정에서 동결융해 후 추가적인 배양 없이 곧바로 핵이식에 사용함으로써 핵이식 과정을 단축시킬 수 있으며, 이를 통해 복제(형질전환 복제) 동물의 생산 효율을 향상시킬 수 있다.
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公开(公告)号：KR1020130050076A
公开(公告)日：2013-05-15
申请号：KR1020110115226
申请日：2011-11-07
Applicant: 대한민국(농촌진흥청장)
IPC: C12N15/873 , C12N5/10
Abstract: PURPOSE: A method for treating frozen and thawed donor cells for improving the productivity of cloned animals is provided to shorten nuclear transplantation without additional culturing. CONSTITUTION: A method for treating frozen and thawed donor cells comprises: a step of culturing somatic cells for synchronization to a G0/G1 phase; a step of quickly freezing the somatic cells at -70 to -120 deg. C; and a step of thawing the somatic cells and transplanting. The somatic cell is transformed.
Abstract translation: 目的：提供一种用于治疗冷冻和解冻供体细胞以提高克隆动物生产力的方法，以缩短核移植而无需额外培养。 构成：用于治疗冷冻和解冻供体细胞的方法包括：将培养体细胞与G0 / G1期同步的步骤; 将体细胞快速冷冻至-70〜-120度的步骤。 C; 并解冻体细胞和移植的一个步骤。 体细胞被转化。
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公开(公告)号：KR101174493B1
公开(公告)日：2012-08-22
申请号：KR1020100038702
申请日：2010-04-26
Applicant: 대한민국(농촌진흥청장)
IPC: A01K67/027 , C12N15/12 , C12N15/85 , C12Q1/68
Abstract: 본발명은제2형당뇨병환자에게서관찰되는 PEA15 유전자의과발현현상을생쥐 PEA15 유전자를사용하여돼지에서과발현되도록한 것으로, 당뇨병치료제개발및 효능검증과병인해석등에필요한전임상당뇨질환모델동물의개발이전단계의것이다.
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公开(公告)号：KR1020070075831A
公开(公告)日：2007-07-24
申请号：KR1020060004508
申请日：2006-01-16
Applicant: 대한민국(농촌진흥청장)
Abstract: A recombinant human von Willebrand Factor(vWF) expression vector is provided to produce the vWF of a human blood coagulation factor through milk of a pig, thereby mass-producing a highly-valued medical protein through cattle. The recombinant human vWF expression vector includes a nucleotide sequence described as SEQ ID : NO. 1 and is constructed by linking a vWF gene of human to the middle of a bovine alpha-S1-casein promoter having a restriction enzyme site Not I and Sal I of a gene for micro-injection and a hGH Poly A. The primer sequence for identifying the introduction of the vWF gene in a genomic DNA of a transgenic pig to which the expression vector is introduced has SEQ ID : NOs. 3-10. The transgenic pig secrets a human vWF agent as milk by introduction of the expression vector thereinto.
Abstract translation: 提供重组人血管性血友病因子（vWF）表达载体以通过猪的牛奶产生人凝血因子的vWF，从而通过牛批量生产高价值的医学蛋白质。 重组人vWF表达载体包含SEQ ID NO： 并且通过将人的vWF基因连接到具有用于微注射的基因的限制性内切酶酶切位点Not I和Sal I以及hGH Poly A的牛α-S1-酪蛋白启动子的中间进行构建。用于 鉴定引入表达载体的转基因猪的基因组DNA中的vWF基因的引入具有SEQ ID：NO。 3-10。 转基因猪通过引入表达载体来分泌人类vWF作为牛奶。
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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。
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