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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Abbreviations
- FPBS:
-
Fibrizyme in pBluscript
- gfp:
-
Green fluorescent protein
- rbs:
-
Ribosomal binding site
- EK:
-
Enterokinase
- SD:
-
Shine-dalgarno
- UTR:
-
Untranslated region
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Acknowledgements
We are grateful to Professor Soon-Chel Park of Chung-Ang University for kindly providing fibrizyme cDNA. This work was supported by the following: a grant from the Crop Functional Genomics Center of the 21st Century Frontier Research Program and the DAEDUK Innopolis of Ministry of Education, Science, and Technology of the Republic of Korea.
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Ko, SM., Yoo, BH., Lim, JM. et al. Production of Fibrinolytic Enzyme in Plastid-Transformed Tobacco Plants. Plant Mol Biol Rep 27, 448–453 (2009). https://doi.org/10.1007/s11105-009-0113-4
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DOI: https://doi.org/10.1007/s11105-009-0113-4