Genetic transformation ofStreptomyces caespitosus |
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Authors: | Jin Cheol Yoo Jung -Bo Sim Sung -Jun Kim Si -Wouk Kim Jung -Jun Lee |
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Institution: | 1. Department of Pharmacy, Chosun University, 501-759, Kwangju, Korea 3. Department of Genetic Engineering, Chosun University, 501-759, Kwangju, Korea 4. Department of Environmental Science, Chosun University, 501-759, Kwangju, Korea 5. Genetic Enginneering Research Institute, KIST, 305-606, Taejeon, Korea
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Abstract: | Genetic transformation ofStreptomyces caespitosus by plasmid pIj 702 was carried out. Optimal conditions for the protoplast preparation ofStreptomyces caespitosus, its regeneration, and its transformation by pIj 702 were evaluated. Addition of 2% glycine to the culture broth was optimal
for protoplast yield. Formation and regeneration of protoplasts were most efficient when the mycelium were harvested at between
late log and stationary growth phase. The regeneration frequency of the protoplasts was 15% when the protoplasts were regenerated
on R2YE agar media containing 0.5M sucrose. Under the best condition for protoplats regeneration, the optimal transformation
frequency was achieved with 40% polyethylene glycol (M.W. 4,000) treatment for 2 minutes. |
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Keywords: | Streptomyces caespitosus Protoplast Regeneration Transformation |
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