^ Donnelly ML, Luke G, Mehrotra A, Li X, Hughes LE, Gani D, Ryan MD (May 2001).^ "Cleavage Activity of Aphtho- and Cardiovirus 2A Oligopeptidic Sequences"."2A self-cleaving peptide-based multi-gene expression system in the silkworm Bombyx mori". ^ Wang Y, Wang F, Wang R, Zhao P, Xia Q (November 2015)."Design and construction of 2A peptide-linked multicistronic vectors". ^ a b Szymczak-Workman AL, Vignali KM, Vignali DA (February 2012)."Cleavage of foot-and-mouth disease virus polyprotein is mediated by residues located within a 19 amino acid sequence". ^ Ryan MD, King AM, Thomas GP (November 1991)."Structures and Corresponding Functions of Five Types of Picornaviral 2A Proteins". ^ Yang X, Cheng A, Wang M, Jia R, Sun K, Pan K, et al."Occurrence, function and evolutionary origins of '2A-like' sequences in virus genomes". ^ Luke GA, de Felipe P, Lukashev A, Kallioinen SE, Bruno EA, Ryan MD (April 2008).Live Imaging In Zebrafish: Insights Into Development And Disease. ^ Sampath Karuna Roy Sudipto (30 August 2010)."Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector". ^ a b c d Liu Z, Chen O, Wall JB, Zheng M, Zhou Y, Wang L, et al.However, the level of drop-off detected in this study varied widely depending on the exact construct used, with some constructs showing little evidence of drop-off furthermore, within a tri-cistronic transcript it reported a higher level of ribosome drop-off after one 2A sequence than after two 2As combined, which is at odds with a linear model of translation. One study reported that 2A sites cause the ribosome to fall off approximately 60% of the time, and that, together with ribosome read-through of about 10% for P2A and T2A, this results in reducing expression of the downstream peptide chain by about 70%. Therefore, up to 50% of F2A-linked proteins can remain in the cell as a fusion protein, which might cause some unpredictable outcomes, including a gain of function. Incomplete cleavage ĭifferent 2A peptides have different efficiencies of self-cleaving, T2A and P2A being the most and F2A the least efficient. ĢA peptides, when combined (or not) with the IRES elements, can make it possible to generate multiple separated peptides within a single transcript. Inserting the CDS of a 2A peptide into the fusing point or replacing the linker sequence with the CDS of a 2A peptide protein can cleave the fused protein into two separated peptides, rescuing the function of the two peptides. The 2A peptides can be applied when the fused protein doesn’t work. In genetic engineering, the 2A peptides are used to cleave a longer peptide into two shorter peptides. However, it is believed to involve ribosomal "skipping" of glycyl-prolyl peptide bond formation rather than true proteolytic cleavage. The exact molecular mechanism of 2A-peptide-mediated cleavage is still unknown. The apparent cleavage is triggered by ribosomal skipping of the peptide bond between the proline (P) and glycine (G) in C-terminal of 2A peptide, resulting in the peptide located upstream of the 2A peptide to have extra amino acids on its C-terminal end while the peptide located downstream the 2A peptide will have an extra proline on its N-terminal end. Adding the optional linker “ G S G” (Gly-Ser-Gly) on the N-terminal of a 2A peptide helps with efficiency. The following table shows the sequences of four members of 2A peptides. F2A is derived from foot-and-mouth disease virus 18 E2A is derived from equine rhinitis A virus P2A is derived from porcine teschovirus-1 2A T2A is derived from thosea asigna virus 2A. Members įour members of 2A peptides family are frequently used in life science research. The name "2A" itself comes from the gene numbering scheme of this virus. For example, F2A, the first described 2A peptide, is derived from foot-and-mouth disease virus. The members of 2A peptides are named after the virus in which they have been first described. 2A peptides are used to help generate polyproteins by causing the ribosome to fail at making a peptide bond. These peptides share a core sequence motif of D圎xNPGP, and are found in a wide range of viral families. Class of peptides An illustration of 2A peptide function: when the CDS of a 2A peptide is inserted between two CDSs of protein, the peptide will be translated into two proteins folding independently due to ribosome skipping (not due to cleavage as shown in the figure).ĢA self-cleaving peptides, or 2A peptides, is a class of 18–22 aa-long peptides, which can induce ribosomal skipping during translation of a protein in a cell.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |