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Functional verification of yeast signal peptide secretion

Service Overview

Nanjing Ruiyuan Biotechnology offers yeast signal peptide secretion verification services. In the experiment, the yeast strain selected was YTK12, which lacks the sucrose conversion enzyme gene and cannot grow on a medium with sucrose as the sole carbon source. The vector plasmid selected contained the tryptophan synthesis gene and a sucrose conversion enzyme gene (SUC2) with a missing signal peptide and the start codon ATG. When a functional signal peptide is linked to the sucrose conversion enzyme gene of pSUC2, the secretion function of the sucrose conversion enzyme can be restored, enabling the YTK12 strain to grow on a defective medium with raffinose as the carbon source. In addition, the TTC color reaction is also commonly used to verify the results.

Technical advantages

1. Facilitates observation: The signal peptide secretion function can be observed directly through color reaction.

2. Stable and efficient: The yeast signal peptide function verification system has been widely applied in biological research at present.

Service Content

Service Content	Weekday (working day)	Delivery Content
Gene synthesis up to pSUC2	15个	1. Construction of the plasmid into the target vector 2. Experimental electronic report 3. Original experimental photos
Transform the yeast cells to the YTK12 yeast strain	7个
Detection of yeast signal peptide tracking	7个
TTC chemical colorimetric detection	7个
Aggregate	36个

Material Sample Processing And Preservation

Type		Sample Submission Standards	Conditions Of Carriage
Choose one from the three options	The sequence to be verified	Our company is engaged in gene synthesis.
	The constructed gene plasmid	Plasmid concentration ≥ 100 ng/μL; Plasmid volume ≥ 30 μL; There is a verification sequencing file	Ice bag
	Genetic plasmid bacterial solution	Each tube volume is ≥ 500 μL; There are identification and sequencing files	Dry ice delivery: Dry ice usage: 1 day (5 kg), 2-3 days (10-20 kg), 3-5 days (20-30 kg).

Case Show

Figure: Utilization ability for disaccharides

References

[1]Xu Q, Tang C, Wang X, Sun S, Zhao J, Kang Z, Wang X. (2019). An effector protein of the wheat stripe rust fungus targets chloroplasts and suppresses chloroplast function. Nat Commun 10, 5571.

[2] Yin W, Wang Y, Chen T, Lin Y, Luo C. (2018). Functional evaluation of the signal peptides of secreted proteins. Bio-protocol 8(9): e2839. DOI: 10.21769/BioProtoc.2839.

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