Red Light Controllable Yeast System

The in-depth characterization of the red light dependent interaction between phytochrome B (PhyB) and phytochrome interaction factor 3 (PIF3) from Arabidopsis allowed the development of the first light switching gene promoter system in yeast. Following this pioneering work, the PhyB-PIF3 system was used to investigate the specific protein-protein interaction mediated by red light in yeast.

Creative BioMart is now able to insert photoreceptors and their interacting proteins to the transcriptional activation elements of target genes so that the transcription can be regulated by red/ far red light.

Construction of the Red Light Controllable Yeast System

Based on PhyB/PIF3 interaction, the system can work as a transfer switch, and the on / off state is controlled by red light / far red light. The system is based on a yeast two hybrid system as a fission transcription factor, in which PhyB is connected to Gal4 DNA binding domain (DBD), and PIF3 is fused to Gal4 trans activation domain (AD). Therefore, under the stimulation of red light (660 nm), the interaction between PhyB and PIF3 reconstructs an active transcription factor that binds to the GAL1 promoter and activates the expression of the target gene.

Creative BioMart has been engaged in microbial gene editing for many years and has a perfect Saccharomyces cerevisiae gene editing technology platform. Our gene editing technology platform has experienced technicians, efficient experimental process and perfect experimental equipment. Our scientists can customize one-to-one solutions according to the specific needs of customers, and provide editing services for a variety of microorganisms to meet the needs of customers.

Recombinant Protein Expression System

Red light can replace chemical inducer to induce the expression of recombinant protein in yeast system. Creative BioMart has been committed to the expression and preparation of recombinant proteins for many years. Our scientists have rich experience in recombinant expression, fermentation and purification of yeast protein, and can quickly customize protein yeast expression strategies for different customers.

Step	Description	Time
Subcloning of yeast expression vector	Codon optimization, gene synthesis (or customer provided plasmid template: target gene amplification) The target gene was subcloned into a suitable expression plasmid Splice sequence validation: sequencing Delivery: correctly constructed recombinant expression plasmid, DH5 containing recombinant plasmid α Strain and sequencing original report	1-2 weeks
Yeast expressing strain transformation	Amplify and culture 5-10 positive clones and induce the expression of recombinant protein SDS-PAGE delivery: report of positive clone strains and expression screening results.	2 weeks
Screening of yeast expressing strains	Detection of recombinant protein expression (applicable to multi copy number and high expression strains) Delivery: report of positive clone strains and expression screening results.	2 weeks
Optimization of yeast expression strains	For the selected monoclonal strains, the culture and induction conditions (medium, cell density, methanol concentration, induction time, etc.) were optimized to improve the expression of the target protein	2-3 weeks
Small scale protein expression and purification	1L fermentation broth was purified by affinity chromatography, ion exchange, hydrophobicity and gel filtration. SDS-PAGE delivery: recombinant protein and purification report. The final protein containing purified tag or excised purified tag can be provided according to customer requirements, with purity > 85%.	2-3 weeks
Large scale protein expression and purification	10L, 30L, 80L, 130L, 500L fermentation scale.

Creative BioMart is always devote us to provide high-quality and satisfactory service to our customers, if you are interested in our services or have some question, please feel free to contact us or make online inquiry.