linkers - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki
linker
Linker#1
L1: GGGGSGGGGS
GGAGGTGGAGGTTCTGGTGGAGGTGGTTCA
Ref?? Rosana
Linker#2
http://parts.igem.org/Protein_domains/Linker
GlyGlyGlyGlyGlySer = GGT GGA GGT GGA GGA TCT
"Several other types of flexible linkers, including KESGSVSSEQLAQFRSLD and EGKSSGSGSESKST"
Chen, Xiaoying, Jennica L. Zaro, and Wei-Chiang Shen. 2013. “Fusion Protein
Linkers: Property, Design and Functionality.” Advanced Drug Delivery Reviews
65 (10) (October): 1357–1369.
EGKSSGSGSESKST 14 aa = 14 * 3 = 42 bp <====<<<< Good size ESGSVSSEQLAQFRSLD 18 aa = 18 * 3 = 54 bp
The two linkers above are from:
Bird, R. E., K. D. Hardman, J. W. Jacobson, S. Johnson, B. M. Kaufman, S. M.
Lee, T. Lee, S. H. Pope, G. S. Riordan, and M. Whitlow. 1988. “Single-Chain
Antigen-Binding Proteins.” Science 242 (4877) (October 21): 423–426.
DNA sequence made: https://www.bioinformatics.org/sms2/rev_trans.html https://www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi?species=4932&aa=1&style=GCG
gaaggtaaatcttctggttctggttctgaatctaaatctact E G K S S G S G S E S K S T
Optimized linker for yEGFP fusion proteins:
GDGAGL (Gly-Asp-Gly-Ala-Gly-Leu)
DNA: ggtgacggtgctggttta
Sheff, M. A., & Thorn, K. S. (2004). Optimized cassettes for fluorescent
protein tagging in Saccharomyces cerevisiae. Yeast , 21(8), 661–670.
Other GFP linker:
GSAGSAAGSGEF (as good as the (GGGGS)4 linker, but less likely for the coding DNA sequence to be removed by homologous recombination)
DNA: GGATCCGCTGGCTCCGCTGCTGGTTCTGGCGAATTC
https://www.nature.com/articles/nbt0799_691
Other info:
More information about fusion protein linkers: https://www.ncbi.nlm.nih.gov/pubmed/23026637
https://www.researchgate.net/post/What_is_the_best_linker_for_a_fusion_protein
There are linkers based on the same principle than the (GGGGS)n which I find superior as they don't have repeats that can lead to homologous recombination. One example: GSAGSAAGSGEF. ref for the GSAGSAAGSGEF linker:
Waldo, G.S., Standish, B.M., Berendzen, J., and Terwilliger, T.C. (1999). Rapid protein- folding assay using green fluorescent protein. Nat. Biotechnol. 17, 691–695.
ref for other similar linkers:
Chen, X., Zaro, J.L., and Shen, W.-C. (2013). Fusion protein linkers: property, design and functionality. Adv. Drug Deliv. Rev. 65, 1357–1369.