国家自然科学基金(30600331), 国家高技术研究发展计划(863)(2007AA100502)和上海市自然科学基金(06ZR14054)资助项目
This work was supported by grants from The National Natural Science Foundation of China (30600331), Hi-Tech Research and Development Program of China(2007AA100502) and Shanghai Natural Science Foundation (06ZR14054)
在NIH3T3细胞中构建了一种链霉菌噬菌体ФC31整合酶报告系统.该报告载体同时编码红色荧光蛋白和绿色荧光蛋白,与编码ФC31整合酶的载体共转染可以反映ФC31整合酶的活性.细胞中从红色荧光到绿色荧光的变化和百分比的变化可经流式细胞仪检出.随着转染中ФC31整合酶表达载体的比例升高,表达绿色荧光的细胞比例上升.ФC31整合酶表达载体和报告系统载体比例在10∶1时,可达最高约90%的红绿荧光转变率.这表明该ФC31整合酶报告系统提供了一种在细胞中快捷可靠的评价ФC31整合酶功能的方法.
A reporter system for ФC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was co-transfected with the plasmid coding ФC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding ФC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-ФC31-integrase]/[reporter plasmid] at 10∶1. This suggests that the ФC31 integrase reporter system provides a probe for the function of ФC31 integrase in cells.
徐焕宇,马晴雯,任兆瑞,巩芷娟,黄淑帧,曾凡一,曾溢滔.一种快捷可靠评价链霉菌噬菌体ФC31整合酶功能的双荧光报告系统[J].生物化学与生物物理进展,2009,36(7):929-933
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