Part:BBa_K3829011
P-ss-yeGFP-V5tag-Anchor protein 5105-T
Our part BBa_K33829011 is a recombinant yeGFP improved from the part reporter GFP BBa_K3402000. We optimized the codon and added a stronger promoter BBa_K3829001 and terminator BBa_K3829000. In our project, yeGFP was used to screen anchored proteins.
Characterization
In our experiment, signal peptide (SS) was used. SS was a short peptide that could secrete proteins outside the cell. We constructed different gene circuits as shown in Figure 1 to verify the function of SS. The results showed that When SS was not present, fluorescence diffused in the cell (positive control). When SS was present (4609 and 5105), the fluorescence was displayed on the cell membrane, indicating that SS could secrete proteins outside the cell.
References
1.Eisenhaber, Birgit, et al. "A sensitive predictor for potential GPI lipid modification sites in fungal protein sequences and its application to genome-wide studies for Aspergillus nidulans, Candida albicans Neurospora crassa, Saccharomyces cerevisiae and Schizosaccharomyces pombe." Journal of molecular biology 337.2 (2004): 243-253.
2.Möller, Steffen, Michael DR Croning, and Rolf Apweiler. "Evaluation of methods for the prediction of membrane spanning regions." Bioinformatics 17.7 (2001): 646-653.
3.Smith MR, Khera E, Wen F. “Engineering Novel and Improved Biocatalysts by Cell Surface Display.” Ind Eng Chem Res, volume 53, issue 16, 29 April 2015, pp. 4021-4032.
4.Tanaka T, Yamada R, Ogino C, Kondo A. “Recent Developments in Yeast Cell Surface Display toward Extended Applications in Biotechnology.” Appl Microbiol Biotechnol, volume 75, issue 3, August 2012, pp. 577-591.
5.Andreu C, Del Olmo ML. “Yeast Arming Systems: pros and cons of different protein anchors and other elements required for display.” Appl Microbiol Biotechnol, volume 102, issue 6, Mar 2018, pp. 2543-2561.
Sequence and Features| None |