Difference between revisions of "Part:BBa K1963004"

Latest revision as of 13:27, 23 October 2016

An sRNA system for down-regulation of E. coli fliC encoding flagellin.

This is a multi-sequence system for small RNA (sRNA) production in an E. coli host - or especially with co-expression of E. coli Hfq BBa_K1963000. The system is based on the BBa_K1963002 template and has the proD strong promoter, followed by a short antisense sequence targeting the 5' end of the fliC transcript, followed by the E. coli micC hairpin, followed by the strong terminator sequence T1/TE. The anti fliC sRNA is placed betweeen C-193 and T-194. The system should impair motility of E. coli.

Usage and Biology

The motile E. coli host strain MG1655 was tranformed with BBa_K1963004 and subjected to a plate-based swimming assay (Figure 1). The BBa_K1963004 contains a sRNA that will target the initial codons of the fliC transcript that encodes flagellin - the principle component of the flagellum. The data related to this biobrick are labelled spiRNA-fliC-CDS in Figure 1.

experimental procedure needed.

Figure 1. sRNA Devices can Inhibit Swimming in an E. coli Test System. Photographs representative of motility assays with MG1655 transformed with BBa_K1963002 (control), BBa_K1963004 (spiRNA-fliC-CDS)and BBa_K1963010 (spiRNA-fliC-RBS-CDS). Bar graph showing average swimming/spreading area and standard deviation of replicates.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 101

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 ===Usage and Biology===
+The motile <i>E. coli</i> host strain MG1655 was tranformed with <partinfo>BBa_K1963004</partinfo> and subjected to a plate-based swimming assay (Figure 1). The <partinfo>BBa_K1963004</partinfo> contains a sRNA that will target the initial codons of the <i>fliC</i> transcript that encodes flagellin - the principle component of the flagellum. The data related to this biobrick are labelled <b><i>spiRNA-fliC-CDS</i></b> in Figure 1.
 experimental procedure needed.
@@ Line 13: / Line 16: @@
 https://static.igem.org/mediawiki/parts/f/f6/Ecoliflicfrulhuq2.png
-Figure 1. Bar graph showing average colony area and standard deviation of replicates. Photographs representative of motility assays with <partinfo>BBa_K1963002</partinfo>, <partinfo>BBa_K1963004</partinfo> and <partinfo>BBa_K1963010</partinfo>.
+<b>Figure 1. sRNA Devices can Inhibit Swimming in an <i>E. coli</i> Test System.</b>
+Photographs representative of motility assays with MG1655 transformed with <partinfo>BBa_K1963002</partinfo> (control), <partinfo>BBa_K1963004</partinfo> (<i>spiRNA-fliC-CDS</i>)and <partinfo>BBa_K1963010</partinfo> (<i>spiRNA-fliC-RBS-CDS</i>). Bar graph showing average swimming/spreading area and standard deviation of replicates.