CD BioSciences offers the luciferase complementation imaging (LCI) service to validate protein-protein interactions. It enables real-time and quantitative analysis in living cells, reflecting proteins' interaction status under plant physiological conditions.

LCI is a new technique developed recently to attach two target proteins to firefly luciferase's C- and N-termini. If the two proteins interact, the N-terminus and C-terminus are close to forming an active luciferase. Based on tobacco transient expression technology and fluorescence imaging equipment, LCI qualifies and quantifies plant protein interactions.

Service Content

Experimental content

• Plant expression vector construction (fusion protein).

a) Cloning or synthesis of the target gene.

b) Restriction endonuclease digestion of skeleton vector and purification.

c) Insertion of target gene into the vector.

d) Transform E. coli, screen positive clones, and sequence for identification.

• Agrobacterium tumefacienstransformation, such as GV3101.
• Inject tobacco.
• Detect fluorescence by plant in vivo imaging system and high sensitivity luminescence detector.
• Quantitative analysis.
• Detect the expression of target proteins.
• Analyze the results and complete the delivery.

LCI Expansion services

• Split-luciferase (split-LCI) complimentary service

Our split-LCI service facilitates dynamic and quantitative in vivo analysis of protein interactions and is a valuable tool for studying protein interaction dynamics by monitoring actin filament breakdown in plants.

• Floated-leaf luciferase complementation imaging (FLuCI) service using binary Gateway vectors

Our service enables rapid and quantitative in vivo analysis of protein interactions following transient expression of split-LUC-tagged interacting proteins, contributing to the quantitative analysis of interactions between large numbers of proteins constituting interaction networks in plant cells.

• Construction service of in vitro LCI mathematical model

To quantitatively understand the relationship between luminescence and protein-pair affinity, we can construct a mathematical model of in vitro LCI based on experimental data, using the Ordinary Differential Equations (ODEs) to represent the entire enzyme reaction system.

• Luciferase screening service

Sample Requirements

Gene template

Sequence Information

Deliverables

• The complete experimental report, including raw data, detailed experimental steps, and collated results (three sets of double luciferase activity pictures).
• Full sequence information and annotation information of the recombinant vector.
• The zymography of the recombinant vector.
• Sequencing results of the recombinant vector.
• Recombinant vector, including one plasmid, one E. coli bacterial fluid, and one Agrobacterium bacterial fluid (25% glycerol bacteria).

Advantages of LCI

• Highly quantitative, allowing linear measurements of luminescence signals over a range of several orders of magnitude.
• In contrast to FRET and BiFC, LCI allows the sampling of entire tissues or cell populations, avoiding bias from individual cells.
• Luminescence is measured in the dark and is not affected by autofluorescence produced from chlorophyll and cell walls.

CD BioSciences specializes in plant protein research. We have cutting-edge facilities and protocols to enable the visualization and characterization of protein interactions in living plant cells. If you have any questions, please contact us and we will help you gain a deeper understanding of plant biology at the molecular level.

For research use only, not for clinical use.