CAJ | 학술논문

将大肠杆菌K-12菌株来源的嘌呤核苷磷酸化酶、尿苷磷酸化酶和胸苷磷酸化酶基因分别克隆到pET-11a载体并转化大肠杆菌BL21(DE3)宿主菌表达,通过SDS-PAGE分析和酶的活性测定,重组菌诱导表达后目的蛋白的表达量占菌体总蛋白的37%以上,与产核苷磷酸化酶的野生菌比较,酶的活性也有显著提高。在特异反应条件下,构建的工程菌可以用来高效催化核苷的转糖基反应。
장대장간균K-12균주래원적표령핵감린산화매、뇨감린산화매화흉감린산화매기인분별극륭도pET-11a재체병전화대장간균BL21(DE3)숙주균표체,통과SDS-PAGE분석화매적활성측정,중조균유도표체후목적단백적표체량점균체총단백적37%이상,여산핵감린산화매적야생균비교,매적활성야유현저제고。재특이반응조건하,구건적공정균가이용래고효최화핵감적전당기반응。
The genes encoding pyrimidine nucleoside phosphorylase,uridine phosphorrylase and thymidine phosphorylase from Escherichia coli K12 were cloned respectively into expression vector pET-11a.The recombinant plasmids were then transformed into the strain E.coli BL21(DE3).As a result the(enzymes) were highly expressed after induction with IPTG.The expression product was analyzed with SDS-PAGE varifying that target proteins were expressed in soluble form and the amount accounts for more than 37% of total protein ...