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Revision as of 13:03, 31 October 2017
BASIC PARTS
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Glycine Glycine Serine (x4) Linker Motif A |
Designed |
· Linker motif used
to provide spacer region · Used to expose
affinity purification tags such as 6x His tag · Serine and glycine
used as they are small and flexible |
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Human Proinsulin Coding Sequence |
Humans |
· Coding sequence for
human proinsulin, sourced from NCIB · Consists of A, B
and C chains, but C-peptide is cleaved out · Contains 3 disulphide bonds · Contains arginine
residues adjacent to C-peptide to allow cleavage |
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Winsulin - Single Chain Insulin Analogue |
Designed |
· Consists of short linker peptide instead of C-peptide · Does not require cleavage therefore simplifies production procedures · Linker peptide raises pI of insulin analogue (long lasting after administration) · Increased thermostability (based on other single chain insulin analogues tested) |
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6x Poly-Histidine Tag codon optimized for use in E. coli |
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· Codon optimized for use in E. coli · Used to immobilize ions on a chromatography affinity column (nickel, cobalt or copper) · Small, and easy to add to the ends of proteins of interest |
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T7 major capsid protein |
· High ribosomal recruitment efficiency · Sourced from the pET15b vectors |
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Ecotin Periplasmic Signal Peptide |
Escherichia coli |
· Small periplasmic, homodimeric protein · Upon fusion to N-terminus of protein will translocate protein to periplasm of E. coli · Periplasm is an oxidative environment ideal for protein folding, and simplifies extraction due to eliminating the need for purifying inclusion bodies from the cytoplasm · Periplasm has reduced proteases |
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YncM Bacillus Secretion Peptide |
Bacillus subtilis |
· Can be fused to the
N-terminus of proteins to induce secretion to the surrounding media wheN proteins are expressed in Bacillus subtilis · Eliminates many
steps in the purification process |
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TEV Protease Cleavage Sequence |
N/A |
· Sequence for which TEV protease is specific to · Can be added between protein of interest and peptides that require removal, such as purification tags |
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Glycine
Glycine Serine (x4) Linker Motif
B |
Designed |
· Linker motif used
to provide spacer region · Used to expose
affinity purification tags such as 6x His tag · Serine and glycine
used as they are small and flexible |