吉林大学学报（医学版）2016，Vol.42Issue(2)：210-214,5.DOI:10.13481/j.1671-587x.20160204
基于FRET技术MMP3生物传感器载体的构建和鉴定
Construction and identification of FRET-based MMP3 biosensor
摘要
Abstract
Objective:To study the construction of matrix metalloproteinase 3 (MMP3)biosensor vector,and to illuminate the activated process of MMP3 in the living cells.Methods:The ECFP-MMP3-YPet biosensor vector anchored on cellular surface was constructed and identified.The MMP3 biosensor was transfected into the 293T cells.The transfection efficiency was observed 24 h after transfection.The flurorescence resonance energy transfer (FRET )-based MMP3 biosensor was observed by inversion fluorescence microscope. Results:The MMP3 biosensor vector was successfully constructed.The length of MMP3-YPet identified by double enzyme digestion and PCR was about 780 bp.The transfection efficiency of MMP3 biosensor was about 40%,and which was evenly presented in cytoplasm of 293T cells.And the FRET ratio of MMP3 biosensor was decreased after stimulation with uPA on the 293T cells. The FRET ratio reached its minimum about 30 min later. Conclusion:The MMP3 biosensor can sensitively and reliably monitor the MMP3 activation in living cells.关键词
荧光共振能量转移/基质金属蛋白酶 3/生物传感器Key words
flurorescence resonance energy transfer/matrix metalloproteinase 3/biosensor分类
医药卫生引用本文复制引用
饶烽,崔钢华,王琰,刘伟,曹薇薇,史晨辉,王维山..基于FRET技术MMP3生物传感器载体的构建和鉴定[J].吉林大学学报（医学版）,2016,42(2):210-214,5.基金项目
国家自然科学基金资助课题（81160225，81260453，81360451）；新疆兵团医药卫生专项资助课题（2013BA020）；兵团国际交流与合作专项基金资助课题（2012BC002，2011BC004）；兵团科技创新团队专项基金资助课题 （）
