This part contains the fim operon under the control of the constitutive promoter P_J23100 (BBa_J23110). The operon consists of coding sequences for expression of six Fim proteins, including FimA, FimI, FimC, FimD, FimF and FimG (Le Trong et al 2010). The constituent genes code for structural proteins, such as FimA which makes up the majority of the main body of a pilus, and others which support the process of pilus biogenesis. When co-transformed with FimH constructs from our project, which have rhamnose-inducible promoters, modified type I pili may be produced. The part requires no induction.
We used a modular cloning strategy to build this composite part from three standard parts (http://parts.igem.org/Part:BBa_K2324016, http://parts.igem.org/Part:BBa_K2324017 and http://parts.igem.org/Part:BBa_K2324018).
Fim Operon expression
The electron micrographs presented above of MG1655 are reminiscent of those produced by Pallesen et. al. and confirm pili production in this strain as expected. Unfortunately when under control of the constitutive P_J23100 promoter pili expression is not seen.
Le Trong, I., Aprikian, P., Kidd, B. A., Thomas, W. E., Sokurenko, E. V., and Stenkamp, R. E. (2010) Donor strand exchange and conformational changes during E. coli fimbrial formation. Journal of Structural Biology 172, 380–388.
PALLESEN, L., POULSEN, L. K., CHRISTIANSEN, G., and KLEMM, P. (1995) Chimeric Fimh Adhesin of Type-1 Fimbriae - a Bacterial Surface Display System for Heterologous Sequences. Microbiology 141, 2839–2848.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 7
Illegal NheI site found at 30
- 21Illegal BamHI site found at 2648
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 931
Illegal AgeI site found at 962
- 1000COMPATIBLE WITH RFC