Part:BBa_K2906102
PepG Keratin Associated Decapeptide w/ HisTag, Type II secB sig, Start & Stop codons
PepG (Cruz et al., 2017b) with START & STOP codons, HisTag and type II secB dependent N-terminal signal sequence. PepG is a small keratin associated peptide which infiltrates the cortex of hair and binds to the keratin structure. Here it can repair damage caused by over-bleaching, restoring strength as well as straightening the hair.
PepG was originally taken from a high-throughput screen of the human genome for keratin associated peptides (Cruz et al., 2017a). PepG would strong associate and would go onto be shown to have restorative effects (Cruz et al., 2017b).
PepG, cystine residues are highlighted in pink
Gel electrophoresis results following primer annealing. N refers to presence of an N-terminal type II secB dependent secretion signal. H refers to the presence of a C-terminal HisTag.
In our experiments we constructed the gene via primer overlaps forming type IIS style overhangs. These could be used via standard type IIS construction from a gBlock if so desired.
Expression was detected via western blot using anti-His antibodies; however, the protein was discovered in cell debris so could be in inclusion bodies.
Left: western blot showing character of decapeptide with HisTag and N-terminal signal sequence. Right: Associated SDS-PAGE
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Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Cruz, C.F., Azoia, N.G., Matamá, T. and Cavaco-Paulo, A. (2017a). Peptide—protein interactions within human hair keratins. International Journal of Biological Macromolecules, 101, pp.805–814.
Cruz, C.F., Martins, M., Egipto, J., Osório, H., Ribeiro, A. and Cavaco-Paulo, A. (2017b). Changing the shape of hair with keratin peptides. RSC Advances, 7(81), pp.51581–51592.
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