Separation of the epitopes in a fusion peptide using flexible linker: Construction, characterization and molecular modeling

The present study aimed to evaluate the effect of flexible linker on solubility, expression level, and folding of fusion peptide containing epitopes of human T lymphotropic virus type ۱. For this purpose, the chimera sequences linked by the flexible linker were inserted into different plasmid vectors and expressed in E. coli strains. The expressed products were analyzed using SDS-PAGE and Western blot techniques. Additionally, the molecular modeling study of the chimera with the flexible linker was performed using iterative threading assembly refinement (I-TASSER) to attain their three-dimensional structures. The chimera expression indicated that the insertion of a flexible (GGGGS)۳ linker among chimera epitopes could induce a high level of expression. The molecular modeling results confirmed our experimental study. This investigation illustrated the key role of linker design in determining the expression level of multi-epitope chimera and conformational folding.

کلیدواژه ها:

Fusion peptide ، HTLV-۱ ، Flexible linker ، Expression level ، Immobilized metal ion affinity chromatography ، Molecular modeling