- 作者： 白壽雄; 莊仁正; 高銘欽; 徐添財
- 作者服務機構： 國防醫學院生物化學系及榮民總醫院醫學研究部
- 中文摘要：
核酸限制?在遺傳工程及DNA圖譜鑑定等研究過程中，扮演極為重要之地位。在已知三百多種核酸限制?中 Eco
R1核酸限制?，可謂用途最廣，已成為核酸研究不可或缺之核酸限制?。
由 E. coli RY13菌種培養，菌體收集及粗製?之製備後，經 Polyethyleneimine 沉澱，硫酸銨沉澱，最後我們使用
Heparin Sepharose-4B 親合力管柱層析純化步驟，此?約純化了42倍，?之比活性可達每毫克蛋白質中含有100,000單
位?活性單位以上。
由上述之純化方法有如下之優點：
一、粗萃?即有相當高之活比性(2,400 U/mg)此部份之?即可作為一般內核酸水解?之應用。
二、整個過程可在兩天內完成且有相當高之回收率(42％)。 - 英文摘要： Restriction endonucleases play a very important role in genetic engineering and DNA mapping.Among hundreds of restriction endonucleases, the Eco R1 enzyme is the most useful and widely investigat-ed enzyme. After sonication and ultracentrifugation, crude extracts of E. coli RY 13 were purified by employingthe polyethyleneimine precipitate, ammonium sulfate precipitate and heparin Sepharose-4B affinity columnchromatography. The Eco R1 enzyme were purified at about 42 folds and the specific activity was about100,000 U/mg of protein. The whole purification procedure was finished within two days. The recovery was about 42%. Theenzyme was sufficiently concentrated for direct specific DNA hydrolysis.
- 中文關鍵字： --
- 英文關鍵字： --
