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Letter 5 -- No Title
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한류의 신화 구조와 문화 이데올로기
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Perinatal outcomes following maternal pre‐exposure prophylaxis (PrEP) use during pregnancy: results from a large PrEP implementation program in Kenya
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Pyrazinamide와 그 대사산물이 유발하는 고뇨산 혈증에 미치는 Pyridoxal의 영향
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On the Calculation and Interpretation of Signal Intensity in Echo-Shifted Sequences
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Unterbrechungsfreie Versorgung
- 검색키워드
- 저자 : 변시명 ( Dong Chul Roh (검색결과 2 건)
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Molecular Cloning of Streptokinase Gene from Streptococcus equisimilis and Its Expression in E. coli
노동출,김종화,박승국,이종우,변시명,Roh, Dong-Chul,Kim, Jong-Hwa,Park, Seung-Kook,Lee, Jong-Woo,Byun, Si-Myung 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.4
The streptococcal genomic DNA digested with Pst I was cloned in E. coli HB101. The overlay technique of casein/plasminogen was used to screen the clones for recombinants carrying the streptokinase gene. The insert size of the plasmid carrying the streptokinase gene was a 2.5, 4.3, and 5.8 Kb, respectively. The restriction maps of all three hybrid plasmids were constructed by digestion with Pst I, Pvu II, Sal I, Hind III, Ava I, BamH I, and Cia I. For the identification of cloned gene, streptokinase was highly purified from S. equisimilis by the methods of gel chromatography and isoelectric focusing and rabbits were immunized with this purified streptokinase. Several lines of evidence, including proof obtained by the immunodiffusion technique, established that the enzyme from E. coli was identical to that from S. equisimilis. In the E. coli cell culture, we found the activity of streptokinase in all three principal locations of the cell. More than 50% were existed in the intracellular space. Streptokinase를 생산하는 균주인 Streptococus equisimilis로 부터 유전자를 분리하고 Pst I으로 분해하여 pBR 322를 vector로 하여 E. coli HB101 대장균에 cloning하였다. Streptokinase를 발현하는 균주를 casein/plasminogen overlay technique를 이용하여 skc를 확인하였다. 이때 2.5 Kb, 4.3 Kb, 5.8 Kb의 DNA 단편이 삽입된 clone이 각각 streptokinase 활성을 보였고 제한 효소 지도를 작성한 결과 2.5 Kb는 4.3 Kb, 5.8 Kb의 일부임을 확인할 수 있었다. 또 streptokinase에 대한 항체를 사용한 immunodiffusion technigue으로 확인한 결과 streptokinase를 생성하는 균주임이 확실하였다. 대장균에서는 streptokinase의 활성이 주요 3부분인 세포내, periplasm, 세포외에 모두 존재 하였고 그 활성은 log phase 후기까지는 배양시간에 따라 비율이 항상 일정하였고, 그 이후에는 periplasm의 활성이 세포외로 나왔으나 세포내에는 50% 이상의 활성이 그대로 존재함을 알 수 있었다.
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Streptococcus equisimillis 로 부터 streptokinae 유전자의 분리와 대장균에서의 발현
노동출,김종화,박승국,이종우,변시명 ( Dong Chul Roh,Jong Hwa Kim,Seung Kook Park,Jong Woo Lee,Si Myung Byun ) 생화학분자생물학회 1986 BMB Reports Vol.19 No.4
The streptococcal genomic DNA digested with Pst I was cloned in E. coli HB101. The overlay technique of casein/plasminogen was used to screen the clones for recombinants carrying the streptokinase gene. The insert size of the plasmid carrying the streptokinase gene was a 2.5, 4.3, and 5.8 Kb, respectively. The restriction maps of all three hybrid plasmids were constructed by digestion with Pst I, Pvu II, Sal I, Hind III, Ava I, BamH I, and Cla I. For the identification of cloned gene, streptokinase was highly purified from S. equisimilis by the methods of gel chromatography and isoelectric focusing and rabbits were immunized with this purified streptokinase. Several lines of evidence, including proof obtained by the immunodiffusion technique, established that the enzyme from E. coli was identical to that from S. equisimilis. In the E. coli cell culture, we found the activity of streptokinase in all three principal locations of the cell. More than 50% were existed in the intracellular space.
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