Production of Fibrinolytic Enzyme in Plastid-Transformed Tobacco Plants |
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Authors: | Suk-Min Ko Byung-Ho Yoo Jong-Min Lim Kwang-Hoon Oh Jae-Il Liu Suk-Weon Kim Jang-Ryol Liu Kwan-Sam Choi Eui-Soo Yoon |
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Institution: | (1) R&D Center, Eugentech Inc., 52 Eoeun-dong, Yuseong-gu, Daejeon, 305-806, Korea;(2) Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 305-806, Korea;(3) Plant Genome Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 305-806, Korea;(4) Department of Applied Biology, Chungnam National University, Daejeon, 305-764, Korea;(5) Department of Biology, Kongju National University, Kongju, 314-701, Korea; |
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Abstract: | The earthworm fibrinolytic enzyme, which belongs to a group of serine proteases with strong fibrinolytic activity, has been
used as an oral drug for prevention and treatment of thrombosis in East Asia. Fibrizyme is a fibrinolytic enzyme isolated
from the earthworm Eisenia andrei. Here we report genetic engineering of tobacco plastids with stable integration of the fibrizyme gene into the tobacco chloroplast
genome. A plastid transformation vector was constructed by introducing various regulatory elements into fibrizyme cDNA. This
vector was delivered by particle bombardment into tobacco leaf explants and plastid-transformed plants were subsequently regenerated
into whole plants through several rounds of selection. We confirmed stable integration of the fibrizyme gene into the tobacco
plastid genome by PCR and Southern blot analyses. Northern and Western blot analyses revealed that mRNA and protein of recombinant
fibrizyme were highly expressed in transformed tobacco plants. |
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