Part:BBa_K2906101
PepG Keratin Associated Decapeptide w/ HisTag & RBS
PepG (Cruz et al., 2017b) with START & STOP codon, RBS and HisTag. 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 tests using western blot with anti-His antibodies could not detect protein in successfully transformed E. coli. We found that E. coli struggle expressing such small peptides from small genes, and based on later discussions with experts we recommend looking into making fusion proteins or repeated polycistronic mRNA.
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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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