원문정보
Solvent Effect on Restriction Endonuclease : Alteration of Specificity of Restriction Endonuclease PvuII in Hydrophobic Solution
초록
영어
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 PvuII 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↓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.
목차
서론
실험재료 및 방법
효소 및 시약
pGEM3의 분리 정제
PvuII의 효소반응
전기영동
PvuII에 미치는 유기용매의 효과
PvuII에 미치는 pH, 반응온도, 반응시간의 효과
PvuII의 절단자리 결정
PvuII 인식자리가 제거된 재조합 벡터 제조
실험 결과
PvuII에 미치는 유기용매의 효과
유기용매에서 특이성에 미치는 pH, 온도, 반응시간의 효과
유기용매에서 PvuII의 절단자리 결정
고찰
결론
감사
참고문헌