YUAN SHUN-ZONG, TAN JIANG-LIN, PENG XU, et al. Interaction between ITGB4BP and P311 measured by fluorescence resonance energy transfer. [J]. Journal of army medical university, 2007, 29(16): 1555-1558.
DOI:
YUAN SHUN-ZONG, TAN JIANG-LIN, PENG XU, et al. Interaction between ITGB4BP and P311 measured by fluorescence resonance energy transfer. [J]. Journal of army medical university, 2007, 29(16): 1555-1558.DOI:
Interaction between ITGB4BP and P311 measured by fluorescence resonance energy transfer
Objective To detect the interaction between ITGB4BP and P311 by fluorescence resonance energy transfer(FRET). Methods Target fragments P311 and ITGB4BP were subcloned into the DsRed expression vector pDsRed1-N1 and pEGFP-N2
respectively. After the recombinants pP311-DsRed and pITGB4BP-EGFP identified by restriction enzyme digestion and sequencing analysis
the 2 constructs were co-transfected into human embryo kidney 293(HEK293)cells
then the energy transfer efficiency (E) and the molecular distances (R) between P311 and ITGB4BP were detected by FRET through selectively photobleaching the acceptors (P311-DsRed). Results Double restriction enzyme digestion showed that the 2 recombinant vectors
pP311-DsRed and pITGB4BP-EGFP were constructed correctly. Both vectors could express in HEK293 cells
and the fluorescence fusion proteins mainly distributed in the cytoplasm and nuclei and were co-localized. E was equal to 14% and R to 6.3 nm in our experiment. Conclusion The recombinant vectors pP311-DsRed and pITGB4BP-EGFP encoding P311-DsRed and ITGB4BP-EGFP distributing in the cytoplasm and nuclei in mammalian cells are constructed successfully
and the interaction between ITGB4BP and P311 can be detected by FRET.
关键词
ITGB4BPP311荧光共振能量转移蛋白相互作用
Keywords
ITGB4BPP311fluorescence resonance energy transferprotein-protein interaction
Department of Neonatology, Key Laboratory of Child Development and Disorders of Ministry of Education, Chongqing Key Laboratory of Pediatrics, Chongqing International Science and Technology Cooperation Center for Child Development and disorders, the Chongqing Children’s Hospital of Chongqing Medical University