• 제목/요약/키워드: a cloned human

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인간 HER-2 재조합 단백질을 사용한 항 HER-2 항체 단백질의 ELISA 정량 방법 개발 (Development of a Quantitative ELISA for Anti HER-2 Antibodies using Human HER-2 Recombinant Proteins)

  • 정선기;류창선;정규진;송규용;김상겸
    • 약학회지
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    • 제55권1호
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    • pp.16-21
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    • 2011
  • HER-2 (Human Epidermal Growth Factor Receptor-2) is a protein giving higher aggressiveness in human breast cancers. Trastuzumab is a monoclonal antibody that targets HER-2 and is known to extend survival across all stages of HER2-positive breast cancer. In this study, we attempted to development of a quantitative ELISA (Enzyme-Linked ImmunoSorbent Assay) for evaluating anti HER-2 antibodies using human HER-2 recombinant proteins to support antibody producing processes and pharmacokinetic studies. We established direct or indirect ELISA method for the trastuzumab-like protein combined human recombinant HER-2. The ELISA method will prove to be great value in quantitating anti-HER-2 antibodies levels for developing anticancer antibodies.

A Rare and Often Unrecognized Brain Meningitis and Hepatopneumonic Congestion are a Major Cause of Sudden Death in Somatic Cloned Piglets

  • 박미령;조성근;임여정;박종주;김진회
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.18-18
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    • 2003
  • In human, sudden infant death syndrome(SIDS) is synonyms for the sudden, unexpected and unexplained death of an infant. The incidence of SIDS has been estimated to be from 1 to 3%. Cloning has a relatively high rate of late abortion and early postnatal death, particularly when somatic cells are used as donors of nuclei and rates as high as 40 to 70% have been reported. However, the mechanisms for SIDS in cloned animals are not known yet. To date, few reports provide detailed information regarding phenotypic abnormality of cloned pigs. In this study, most of the cloned piglets were alive at term and readily recovered respiration. However, approximately 82% of male cloned piglets (81/22) died within a week after birth. Significant findings from histological examinations showed that 42% of somatic cloned male piglets died earlier than somatic cloned female piglets, most probably due to severe congestion of lung and liver or neutrophilic inflammation in brain, which indicates that unexpected phenotypes can appear as a result of somatic cell cloning. No anatomical defects in cloned female piglets were detected, but three of the piglets had died by diarrhea due to bacterial infection within 15 days after birth. Although most of male cloned piglets can be born normal in terms of gross anatomy, they develop phenotypic anomalies that include leydig cell hypoplasia and growth retardation post-delivery under adverse fetal environment and depigmentation of hair- and skin-color form puberty onset. This may provide a mechanism for development of multiple organ system failure in some cloned piglets. Th birth weights of male cloned pig in comparison with those of female cloned piglets are significantly reduced(0.8 vs 1.4kg) and showed longer gestational day(120 vs 114). In conclusion, brain meningitis and hepatopneumonic congestion are a major risk factor for SIDS and such pregnancy in cloned animals requires close and intensive antenatal monitoring.

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<네버렛미고>를 통해본 복제 인간 윤리 (Ethics for Cloned Human Beings: )

  • 김미혜
    • 한국콘텐츠학회논문지
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    • 제17권8호
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    • pp.121-129
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    • 2017
  • 생명공학의 발달로 인해 복제 인간을 만들기 위해 인간의 유전자를 복제한다는 가상의 시나리오는 더 이상 낯설지 않다. <네버렛미고>의 등장인물들은 100세 수명기획에 의해 실험실에서 만들어진 복제 인간들이다. 이들은 해일샴이라는 학교에서 공동생활을 하면서 은밀하게 사육되는데, 이 프로젝트의 목적은 불치병에 걸린 진짜 인간 환자들에게 건강한 장기를 제공하기 위한 것이다. 주인공 캐시, 토미, 그리고 루스는 사춘기 시절 이곳에서 육체와 의식의 성장을 경험하며 자신들이 복제 인간이라는 정체성의 비밀도 알게 된다. 성인이 되어 이들은 두 번째 거주지 코티지로 이동하여 장기기증을 시작할 준비를 한다. 두 번째 단계 또한 좀 더 진짜 성인 인간의 장기와 유사한 장기를 만들어 제공하기 위한 프로그램의 일환이다. 인간들이 꾸민 모든 계획을 다 알고 있지만, 이들은 그것에 대해 저항하지 않고 자신들이 처한 상황을 숙명적으로 수용한다. 그러나 이들의 무저항은 삶에 대한 포기 선언이 아니라 자신들의 장기 기증을 통해 생명의 연장이라는 또 다른 미래를 위한 자기희생적 생명 연장이다. 영화는 복제된 인간들의 우애와 희생적 태도를 강조해서 보여줌으로써 난치병의 치료를 위한 생명공학과 생명윤리라는 상이한 견해에 대해 철학적 사유가 뒷받침된 생명윤리적 관점에서 복제 인간에 대한 논의를 이어갈 필요가 있음을 보여준다.

Pharmacogenomic Application for Gene Targeting and Molecular Characterization of a Nucleoside Transporter hCNT2 in Human Intestine

  • Shin, Ho-Chul;Lee, Jong-Hwa;Lee, Han-Ok;Duxin Sun;Gordon L. Amidon
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2003년도 추계학술대회
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    • pp.158-159
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    • 2003
  • We have cloned and functionally expressed a sodium dependent human nucleoside transporter, hCNT2, from a CNS cancer cell line U251. Our cDNA clone of hCNT2 had the same predicted amino acid sequence as the previously cloned hCNT2 transporter. Of the several cell lines studied, the best hCNT2 transport function was obtained when transiently expressed in U251 cells.(omitted)

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Metabolism of Rutin and Poncirin by Human Intestinal Microbiota and Cloning of Their Metabolizing α-L-Rhamnosidase from Bifidobacterium dentium

  • Bang, Seo-Hyeon;Hyun, Yang-Jin;Shim, Juwon;Hong, Sung-Woon;Kim, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제25권1호
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    • pp.18-25
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    • 2015
  • To understand the metabolism of flavonoid rhamnoglycosides by human intestinal microbiota, we measured the metabolic activity of rutin and poncirin (distributed in many functional foods and herbal medicine) by 100 human stool specimens. The average α-L-rhamnosidase activities on the p-nitrophenyl-α-L-rhamnopyranoside, rutin, and poncirin subtrates were 0.10 ± 0.07, 0.25 ± 0.08, and 0.15 ± 0.09 pmol/min/mg, respectively. To investigate the enzymatic properties, α-L-rhamnosidase-producing bacteria were isolated from the specimens, and the α-L-rhamnosidase gene was cloned from a selected organism, Bifidobacterium dentium, and expressed in E. coli. The cloned α-L-rhamnosidase gene contained a 2,673 bp sequcence encoding 890 amino acid residues. The cloned gene was expressed using the pET 26b(+) vector in E. coli BL21, and the expressed enzyme was purified using Ni2+-NTA and Q-HP column chromatography. The specific activity of the purified α-L-rhamnosidase was 23.3 µmol/min/mg. Of the tested natural product constituents, the cloned α-L-rhamnosidase hydrolyzed rutin most potently, followed by poncirin, naringin, and ginsenoside Re. However, it was unable to hydrolyze quercitrin. This is the first report describing the cloning, expression, and characterization of α-L-rhamnosidase, a flavonoid rhamnoglycosidemetabolizing enzyme, from bifidobacteria. Based on these findings, the α-L-rhamnosidase of intestinal bacteria such as B. dentium seem to be more effective in hydrolyzing (1 →6) bonds than (1 →2) bonds of rhamnoglycosides, and may play an important role in the metabolism and pharmacological effect of rhamnoglycosides.

Gene Cloning and Nucleotide Sequence of Human Dihydrolipoamide Dehydrogenase-Binding Protein

  • Lee, Jeongmin;Ryou, Chongsuk;Jeon, Bong Kyun;Lee, Poongyeon;Woo, Hee-Jong;Kwon, Moosik
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권3호
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    • pp.421-426
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    • 2002
  • The pyruvate dehydrogenase complex (PDC), a member of $\alpha$-keto acid dehydrogenase complex, catalyzes the oxidative decarboxylation of pyruvate with the formation of $CO_2$, acetyl-CoA, NADH, and $H^+$. This complex contains multiple copies of three catalytic components including pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide dehydrogenase (E3). Two regulatory components (E1-kinase and phospho-E1 phosphatase) and functionally less-understood protein (protein X, E3BP) are also involved in the formation of the complex. In this study, we have partially cloned the gene for E3BP in human. Nine putative clones were isolated by human genomic library screening with 1.35 kb fragment of E3BP cDNA as a probe. For investigation of cloned genes, Southern blot analysis and the construction of the restriction map were performed. One of the isolated clones, E3BP741, has a 3 kb-SacI fragment, which contains 200 bp region matched with E3BP cDNA sequences. The matched DNA sequence encodes the carboxyl-terminal portion of lipoyl-bearing domain and hinge region of human E3BP. Differences between yeast E3BP and mammalian E3BP coupled with the remarkable similarity between mammalian E2 and mammalian E3BP were confirmed from the comparison of the nucleotide sequence and the deduced amino acid sequence in the cloned E3BP. Cloning of human E3BP gene and analysis of the gene structure will facilitate the understanding of the role(s) of E3BP in mammalian PDC.

Advancement and Application of Somatic Cell Nuclear Transfer Technique in Dog

  • Oh, H.J.;Hong, S.G.;Park, J.E.;Kim, M.J.;Gomez, M.N.;Kim, M.K.;Kang, J.T.;Kim, J.E.;Jang, G.;Lee, B.C.
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2009년도 특별 Symposium
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    • pp.49-57
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    • 2009
  • The cloning of canids was succeeded in 2005, several years after the birth of Dolly the sheep and also after the cloning of numerous other laboratory and farm animal species. The delay of successful somatic cell nuclear transfer (SCNT)was due to the unique reproductive characteristics of the female dogin comparison to other domestic mammals, such as ovulation of immature canine oocyte and a requirement of 25 days for the completion of meiosis within the oviduct (Holst & Phemister, 1971). When the technology for the recovery of in vivo matured oocyte was established, the application of cloning also became possible and cloned dog offspring were obtained. This report summarizes the progress of technical procedures that are required for cloning canids and the application of this technique. The first cloned dog, Snuppy, was achieved using an in vivo-matured oocyte which was enucleated and transferred with an adult skin cell of male Afghan hound. After establishment of a criterion of well-matured oocyte for the improvement of SCNT efficiency, we obtained three cloned female Afghan hound and a toy poodle cloned from 14 year-old aged Poodle using SCNT through this factor. To date, cloned dogs appeared to be normal and those that have reached puberty have been confirmed to be fertile. Through application of canine SCNT technique, first, we demonstrated that SNCT is useful for conserving the breed of endangered animal from extinction through cloning of endangered gray wolves using inter-species SCNT and keeping the pure pedigree through the cloning of Sapsaree, a Korean natural monument. Secondly, we showed possibility of human disease model cloned dog and transgenic cloned dog production through cloning of red fluorescent protein expressing dog. Finally, SCNT can be used for the propagation of valuable genotypes for making elite seed stock and pet dog. In summary, dog cloning is a reproducible technique that offers the opportunity to preserve valuable genetics and a potential step towards the production of gene targeted transgenic cloned dogs for the study of human diseases.

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Recent Advances in Cloning Technology in the Pig -Review-

  • Miyoshi, K.;Sato, E.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권2호
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    • pp.258-264
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    • 2000
  • Production systems for cloned pigs are very important not only for an increase in production of superior animals but also for the production of knockout animals with organs that do not contain antigens for xenotransplantation or to analyze functions of isolated human genes. At present, however, effective systems have not been developed. We have tried to produce cloned pigs by transfering cultured cells into enucleated oocytes and obtained some cloned embryos. To develop a production system for cloned pigs, the basic technologies needed to support such an effort must be improved.

사람 장내세균군집 유래 Bacteorides stericoris HJ-15의 Chondroitinase ABC의 클로닝 (Cloning of Chondroitinase ABC from Bacteroides stercoris HJ-15, a Human Intestinal Anaerobic Bacterium)

  • 방서현;심주원;현양진;김동현
    • 한국미생물·생명공학회지
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    • 제44권2호
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    • pp.140-144
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    • 2016
  • 사람의 분변으로부터 분리한 Bacteroides stericoris HJ-15로부터 chondroitinase ABC 유전자를 클로닝하였다. 클로닝한 chondroitinase ABC 유전자는 3,090 bp, 1,029 아미노산으로 구성되어 있었다. B. stercoris chondroitinase ABC 유전자는 이미 보고된 chondroitinase ABC 유전자들과 호몰로지가 없었으나, 아미노산서열에서는 82% 호몰로지를 보였다. T7 promoter를 가진 pET-26b+ expression vector에 클로닝한 chondroitinase ABC 유전자를 Escherichia coli BL21 (DE3)에서 발현하여 정제한 재조합 chondroitinase ABC는 chondroitin sulfate A, B 및 C를 모두 분해하였다.