Part:BBa_K204500
FliH
FliH the regulator of ATPase FliI, is one of soluble components of export apparatus. In the absence of FliI, FliH inhibits the protein translocation [1]. Further more, even in the presence of FliI, FliH expression from pTrc99A vector caused pronounced inhibition of swariming motility of the cell [2].
By over expressing FliH, flagellar protein translocation will be inhibited as a result swarming motility will decrease.
To characterize the effect of FliH on the swarming motility and protein secretion level, we used p(Tet) and p(T7) promoters. Salmonella typhimurium (MMHI0117) was transformed with pTet FliH (K204502), pT7_FliH (K204503), and pUC19 for vector control. In this experiment, we used MMHI0117, which is the fliH fliI null mutant. So it was speculated that less FliH expression will be required for wild type cell to control protein translocation. So as first step, we tested our parts in this strain. FliH expression from both parts decreased the swarming motility of the cell. Because T7 promoter is leaky in the absence of T7 RNAP in Salmonella typhimurium, the swarming motility was decreased transforming pT7_FliH (K204503). And the secretion level of flagellar protein is also decreased.
So, these results showed that FliH can be used to control the swarming motility and the secretion level of flagellar protein as we expected. For application for other strain, the check of compatibility and improving FliH expression will be needed.</p>
[1] Minamino, T. & Namba, K. Distinct roles of the FliI ATPase and proton motive force in bacterial flagellar protein export. Nature 451, 485-488 (2008).
[2] Minamino, T. & Macnab, R.M. Interactions among components of the Salmonella flagellar export apparatus and its substrates. Molecular Microbiology 35, 1052-1064 (2000).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 426
- 1000COMPATIBLE WITH RFC[1000]
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