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동물 및 식물세포공학

Production of human factor X in transgenic rice cell suspension culture

초록

영어

Factor X, a vitamin K-dependent blood coagulation glycoprotein, is the precursor for factor Xa, the enzyme component of the prothrombinase complex. The factor X is cleaved during coagulation by several different protease including the intrinsic Xase complex, the factor Xactivating enzyme from Russell's viper venom(RVV) and trypsin, and also by extrinsic pathway to give an active enzyme factor Xa. Factor Xa is often used to remove fusion tags from expressed proteins. Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile-
(Glu or Asp)-Gly-Arg. Human factor X was produced in transgenic rice cell suspension culture with the rice amylase 3D promoter. RAmy3D promoter is highly expressed by sugar starvation. The expression of human factor X as mRNA and protein in transgenic rice cell was detected Northern and Western blot analysis. The enzyme activity of RVV activated human factor X was generally characterized by using EGFP-hTNFa fusion protein as substrate, which contained a
human factor Xa cleavage site between two moieties. These results demonstrate the use of human factor Xa derived from plant cell suspension culture as proteases (This work was supported by the grant from Korean Ministry of Knowledge Economy)..

저자정보

  • Eun-Sun JEUNG Jeonju Biomaterials Institute
  • Yun-Ji SHIN Jeonju Biomaterials Institute
  • Hyo Boon KIM Jeonju Biomaterials Institute
  • Min-Woo NAM Jeonju Biomaterials Institute
  • Hong-Soo DOO Jeonju Biomaterials Institute
  • Tae-Ho KWON Jeonju Biomaterials Institute
  • Moon-Sik YANG Division of Biological Sciences, Chonbuk National University

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