【摘要】 选用Invitrogen公司开发的重组肠激酶,在不同反应条件下消化融合蛋白Trx-Exendin-4,用16%的Tricine-SDS-PAGE检测目的多肽Exendin-4与其他切割产物的量.结果表明:温度对肠激酶消化融合蛋白Trx-Exendin-4的影响很大,37℃时酶活力高,以非特异性切割为主;16℃时酶活力居中,特异性切割与非特异性切割共存;4℃时酶活力较低,但以特异性切割为主.获得Exendin-4的最佳消化条件是:1 U肠激酶在4℃条件下6 h消化3 mg的Trx-Exendin-4,效率最高,特异性最好.更多还原

【Abstract】 The fusion protein Trx-Exendin-4 was digested by using the recombinant enterokinase of Invitrogen in different reaction conditions,then the peptide Exendin-4 and other product of nonspecific cutting were detected with 16% Tricine-SDS-PAGE.The result showed that temperature had large effects on the enterokinase digestion on recombinant protein Trx-Exendin-4.When digesting at 37 ℃,enterokinase had the highest activity without specificity;when digesting at 16 ℃,it had the moderate activity and specificity;when digesting at 4 ℃,it had the lowest activity but highest specificity.The optimal digestion condition for enterokinase was that one unit of enterokinase digested 3 mg of Trx-Exendin-4 at 4 ℃ for 6 h,which had the highest efficiency and specificity.更多还原