Team:Utah State/Project

Antimicrobial spider silk proteins (functionalized spider silk) could be used to make antimicrobial spider silk fibers or films. A previous study by Gomes et al. 2011 demonstrated that it is feasible to produce antimicrobial spider silk films. The Utah State iGEM team in 2012 was able to successfully produce spider silk proteins in E. coli with the use of BioBricks and successfully spin spider silk protein that was approximately 25.4 kDa in size. The parts that were submitted to the registry by Utah State in 2012 are RFC 23 compatible which allows for protein fusions, hence the addition of RFC compatible AMPs to spider silk protein at the C or N terminal would be highly desirable.

The spider silk generator (BBa_K844016) created by Utah State in 2012 was able to produce proteins of 25.4 kDa. An increase of spider silk subunit repeats at the genetic level would enable a larger (hence ‘stronger’ fiber) to be produced. This year we proposed to double the number of repeats to 8 and attach AMPs to either the C or N terminal ends of the spidersilk. The purification of this antimicrobial functionalized silk would be purified with a 10x His Tag, with the His Tag at the opposite end of where the AMP would be located. The figures below demonstrate the design of antimicrobial functionalized spider silk.

All constructs were assembled in pSB1C3 an E. coli as the chassis. AMPs were designed with RFC 23 under consideration to allow for protein fusions.