The chemiluminescence (CL) of luminol-Cu(Ⅱ) was applied to HPLC determination o f ascorbic acid, which was separated by a C 18 reverse-phase column with a mobile phase of 0.25 mol/L HAc. The eluted ascorbic aci...The chemiluminescence (CL) of luminol-Cu(Ⅱ) was applied to HPLC determination o f ascorbic acid, which was separated by a C 18 reverse-phase column with a mobile phase of 0.25 mol/L HAc. The eluted ascorbic acid was mixed with 0.3 mmol/L l um inol and 0.05 mol/L CuSO 4. The light emission from the reaction of Cu(Ⅱ) oxi diz ed ascorbic acid and luminol was detected by a modified luminometer. The detecti on limit was 3.6×10 -6 mol/L for ascorbic acid at a S/N ratio of 3, and the linea r calibration range was 2×10 -4\| 2×10 -3 mol/L. The relative standard deviation for 5 replicate injections of 1×10 -3 mol/L ascorbic acid was calculated as 4.3 % . The method was successfully applied to determination of ascorbic acid in juice beverage.展开更多
目的:构建以mLumin荧光蛋白为指示系统的小鼠T细胞免疫球蛋白和结构域蛋白(mouse T cell Ig and ITIM domin,mTIGIT)胞外区真核表达载体,并表达mTIGIT-mLumin融合蛋白。方法:分别设计mTIGIT和mLumin的特异性引物,通过PCR技术扩增两个目...目的:构建以mLumin荧光蛋白为指示系统的小鼠T细胞免疫球蛋白和结构域蛋白(mouse T cell Ig and ITIM domin,mTIGIT)胞外区真核表达载体,并表达mTIGIT-mLumin融合蛋白。方法:分别设计mTIGIT和mLumin的特异性引物,通过PCR技术扩增两个目的基因序列,经酶切、连接,克隆至plenti-puro和pcDNA3.1载体。将构建的plenti-puro-mTIGIT-mLumin慢病毒载体进行病毒包装并感染HEK-293T细胞,而pcDNA3.1-mTIGIT-mLumin真核表达质粒转染非洲绿猴肾成纤维Cos7细胞,利用两种表达载体的药物抗性进行筛选感染或转染的细胞,检测融合蛋白的表达情况。结果:通过PCR顺利扩增获得mTIGIT和mLumin,经酶切、连接插入至载体,基因测序结果证实克隆的mTIGIT和mLumin片段序列正确,并成功正确插入到plenti-puro载体和pcDNA3.1载体中;荧光显微镜和共聚焦显微镜观察显示,转染的HEK-293T细胞和Cos7细胞经过筛选能够稳定表达mTIGIH-mLumin融合蛋白。结论:成功构建plenti-puro-mTIGIT-mLumin慢病毒载体和pcDNA3.1-mTIGIT-mLumin真核表达载体,且能够在目的细胞中稳定表达mTIGIT-mLumin融合蛋白。展开更多
基金National Natural Science Foundation of China(2 96 0 5 0 0 1)
文摘The chemiluminescence (CL) of luminol-Cu(Ⅱ) was applied to HPLC determination o f ascorbic acid, which was separated by a C 18 reverse-phase column with a mobile phase of 0.25 mol/L HAc. The eluted ascorbic acid was mixed with 0.3 mmol/L l um inol and 0.05 mol/L CuSO 4. The light emission from the reaction of Cu(Ⅱ) oxi diz ed ascorbic acid and luminol was detected by a modified luminometer. The detecti on limit was 3.6×10 -6 mol/L for ascorbic acid at a S/N ratio of 3, and the linea r calibration range was 2×10 -4\| 2×10 -3 mol/L. The relative standard deviation for 5 replicate injections of 1×10 -3 mol/L ascorbic acid was calculated as 4.3 % . The method was successfully applied to determination of ascorbic acid in juice beverage.
文摘目的:构建以mLumin荧光蛋白为指示系统的小鼠T细胞免疫球蛋白和结构域蛋白(mouse T cell Ig and ITIM domin,mTIGIT)胞外区真核表达载体,并表达mTIGIT-mLumin融合蛋白。方法:分别设计mTIGIT和mLumin的特异性引物,通过PCR技术扩增两个目的基因序列,经酶切、连接,克隆至plenti-puro和pcDNA3.1载体。将构建的plenti-puro-mTIGIT-mLumin慢病毒载体进行病毒包装并感染HEK-293T细胞,而pcDNA3.1-mTIGIT-mLumin真核表达质粒转染非洲绿猴肾成纤维Cos7细胞,利用两种表达载体的药物抗性进行筛选感染或转染的细胞,检测融合蛋白的表达情况。结果:通过PCR顺利扩增获得mTIGIT和mLumin,经酶切、连接插入至载体,基因测序结果证实克隆的mTIGIT和mLumin片段序列正确,并成功正确插入到plenti-puro载体和pcDNA3.1载体中;荧光显微镜和共聚焦显微镜观察显示,转染的HEK-293T细胞和Cos7细胞经过筛选能够稳定表达mTIGIH-mLumin融合蛋白。结论:成功构建plenti-puro-mTIGIT-mLumin慢病毒载体和pcDNA3.1-mTIGIT-mLumin真核表达载体,且能够在目的细胞中稳定表达mTIGIT-mLumin融合蛋白。
