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KMID : 0380019940090010063
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Korean Journal of Biotechnology and Bioengineering
1994 Volume.9 No. 1 p.63 ~ p.71
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Solvent Effect on Restriction Endonuclease : Alteration of Specificity of Restriction Endonuclease Pvu Ⅱ in Hydrophobic Solution
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Abstract
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During the last decade enzyme reaction in organic solvent has been studied to show that specificity in buffer is different from that in organic solvent. The specificity of restriction enzyme was effected by various factors such as ionic strength, salt, organic solvent and temperature. In this study, restriction enzyme PvuIl which is used most frequently in genetic engineering and the substrate was vector pGEM3 whose sequence was already known were used. As a result the recognition sequence site was changed in the presence of organic solvents whose Log P are -1.5- 0. Their specificities were contrast with activities were contrasted. Specificities were not changed in organic solvent easily in inactivating enzyme. We think that the enzyme recognition site was not changed randomly but by preferential order. A recombinant vector which does not contain typical cleavage site CAG I CTG was cleaved in 20% ethanol solution. This result might show that restriction enzyme could be used to cleave at unusual sites by changing the reaction conditions.
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