Yeast Signal Peptide Tracking and Screening Service

Yeast signal peptide screening technology (YSST) was used to verify the secretion function of signal peptides. The yeast strain selected in the experiment was YTK12 that lacked the sucrose invertase gene and could not grow on the medium with sucrose as the sole carbon source. The selected vector plasmid is pSUC2 containing a tryptophan synthesis gene and a sucrose invertase gene (SUC2) lacking signal peptide and initiation codon ATG. When a functional signal peptide is connected to the front of the sucrose invertase gene of pSUC2, the secretory function of sucrose invertase can be restored, so that the YTK12 strain can grow on a defective medium with raffinose as a carbon source. In addition, TTC color reaction is often used to verify the results.

Service introduction

Creative BioMart provides yeast signal peptide screening services based on years of rich experience in yeast protein expression system research.

Experimental steps

1. Use SignalP to predict the signal peptide region of the candidate secreted protein, design primers, and construct it on the pSUC2 vector.

2. Streak on the VPDA medium to cultivate the YTK12 strain. The constructed expression vector was introduced into YTK12, and spread on the CMD-W (deficient amino acid) medium plate for cultivation.

3. Pick the positive clones and culture them on YPRAA medium with raffinose as the carbon source. If the transferred yeast cells can grow on YPRAA medium, then it proves that the signal peptide has secretory function; if it cannot grow, it proves that the signal peptide has no secretory function.

4. Cultivate the positive transformants in the above steps with CMD-W medium. Collect the cell suspension and centrifuge to remove the supernatant, resuspend the pellet in the resuspension, and incubate at 37°C for 10 min. After centrifugation, take the supernatant into a test tube and add TTC. Take the untransformed YTK12 yeast as a control, observe the color change, and record it with pictures. If the color turns red, it proves that the signal peptide has a secretory function.

Material requirements

• Name and Gene ID of the protein to be studied or plasmid
• Cell sample > 1*10⁷
• Animal sample > 1 g
• Plant samples > 2 g
• Total RNA > 500 μg

Delivery result

• Original picture
• Plasmid and bacterial solution
• Experiment report, including experimental equipment, reagents, methods, results, conclusions.

Project cycle

It takes about 20 working days to complete yeast signal peptide tracking and screening service.

Service process

Please let us know the details of your yeast related projects through online consultation. We will reply to you in time and provide you with a comprehensive evaluation report on your project, including the feasibility evaluation of the project and the detailed experimental plan designed by our professionals. As our condition optimization is completed, the result report and other final products will be delivered to you immediately.

Creative BioMart has accumulated many years of experience in the field of yeast research, helping our customers accelerate research in all fields using yeast as a research tool, and improve the overall success rate of the project. Please tell us your project requirements, and we will provide you with a full service from strategy design to final report. If you have any questions, please feel free to contact us.