An Expedient Reliable Double Fluorescent Reporter System for φC31 Integrase Function Evaluation
10.3724/SP.J.1206.2008.00698
- VernacularTitle:一种快捷可靠评价链霉菌噬菌体φC31整合酶功能的双荧光报告系统
- Author:
Huanyu XU
;
Qingwen MA
;
Zhaorui REN
;
Zhijuan GONG
;
Shuzhen HUANG
;
Fanyi ZENG
;
Yitao ZENG
- Publication Type:Journal Article
- Keywords:
φC31 integrase;
reporter system;
sequence-directed recombination;
NIH3T3 cells
- From:
Progress in Biochemistry and Biophysics
2009;36(7):929-933
- CountryChina
- Language:Chinese
-
Abstract:
A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 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 φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.
