CAJ | 학술논문

利用PCR扩增猪链球菌2型强毒株SS2-1丝氨酸/苏氨酸磷酸酶( STP)的编码基因,克隆到表达载体pET-30a(+),在大肠杆菌BL21中进行原核表达,获得重组蛋白质rSsSTP。重组蛋白质经His蛋白质层析柱纯化后,利用pNPP分析rSsSTP的酶活性。结果显示rSsSTP为Mn2+依赖的磷酸酶,最适离子浓度为2 mmol/L,最佳反应温度为37℃,最佳pH值为8.0。
이용PCR확증저련구균2형강독주SS2-1사안산/소안산린산매( STP)적편마기인,극륭도표체재체pET-30a(+),재대장간균BL21중진행원핵표체,획득중조단백질rSsSTP。중조단백질경His단백질층석주순화후,이용pNPP분석rSsSTP적매활성。결과현시rSsSTP위Mn2+의뢰적린산매,최괄리자농도위2 mmol/L,최가반응온도위37℃,최가pH치위8.0。
In this study, the complete serine/threonine phosphatase ( STP) gene was amplified by PCR from the ge-nome of virulent strain SS2-1 of Streptococcus suis type 2, and cloned into expresssion vector pET-30a(+). The recombi-nant SsSTP protein ( rSsSTP) was induced by IPTG to express in E. coli BL21 and purified using HisTrap columns. The ac-tivity of rSsSTP was tested using p-nitrophenyl phosphate ( pNPP) as a substrate. rSsSTP was a manganese-dependent phos-phatase in the presence of Mn2+at 2 mmol/L concentration; the optimal temperature was 37 ℃,and the optimal pH value was 8. 0.