Difference between revisions of "Part:BBa K1321302"

Revision as of 01:59, 1 November 2014

pSEVA331-BB with J23100-RFP

This part contains Anderson promoter J23117 and RFP coding device in pSEVA331-BB backbone. pSEVA331-BB is a broad host range plasmid converted into biobrick format, and is capable of replication in E.coli and several other G- bacterial species. We have verified that it also replicates in the cellulose producing bacterium Gluconacetobacter xylinus. BBa_K1321302 is a member of the G.xylinus genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332). BBa_J23100 drives a high level of expression in G.xylinus.

Figure 1. RFP expression from BBa_K1321302 in G.xylinus igem strain. Top three plates - transformed G.xylinus, bottom - untransformed control. Image taken under blue light box 5 days after transformation. G.xylinus was cultured on HS-agar plates, at 30degC inverted.

G.xylinus toolkit was designed to ease genetic engineering of cellulose-based biomaterials using the cellulose synthesizing bacterium Gluconacetobacter xylinus (parts Bba_K1321305 and BBa_K1321306). As no registry parts had been tested in G.xylinus, the aim of this toolkit was to determine the parts usable in G.xylinus and to characterize them in this host. pSEVA331-Bb is a non-standard broad host range plasmid capable of replication in G.xylinus and E.coli (a shuttle vector) and was selected because the registry's standard plasmid backbone pSB1C3 can not be used for G.xylinus engineering.

NOTE: Because the registry's standard plasmid backbone pSB1C3 is not capable of replication in Gluconacetobacter species, the G.xylinus genetic engineering toolkit is housed mainly in pSEVA331-Bb. pSEVA331-Bb is a non-standard backbone, which therefore can't be quality controlled by and maintained in the Registry. However, in order to make the G.xylinus toolkit available for the synthetic biology community, Imperial iGEM 2014 team has made it freely available upon request, with quality control provided (see Experience). To request, please contact Imperial iGEM 2014 team.

Sequence and Features