Difference between revisions of "Part:BBa K1963009"

Latest revision as of 22:44, 22 October 2016

Device for rhamnose-dependent expression of an OsmY fusion to Serratia Hfq.

A rhamnose-regulated device based on existing and new (2016) biobricks. This is designed to induce production of a fusion protein between E. coli OsmY (a secerted protein) and Serratia marcescens Hfq (an RNA binding protein). Co-expression with Serratia sRNA may generate a specific RNA secretion system.

Usage and Biology

Experimental procedures here.

Figure 1. Whole cell blots expressing OsmY-Hfq fusion protein under the control of the rhamnose promoter.

Figure 2. Supernatant blots of OsmY-HA (BBa_K1963007), OsmY-Hfq (E. coli- BBa_K1963008) and OsmY-Hfq (Serratia BBa_K1963009).. Western blotting with anti-HA antibody shows secretion of OsmY-HA, and OsmY-Hfq (E. coli) and anti-RNAP antibody shows no antigen detected in the supernatant therefore the cells remained intact and there was no cell lysis.

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
COMPATIBLE WITH RFC[1000]

@@ Line 6: / Line 6: @@
-Usage and Biology
+===Usage and Biology===
+Experimental procedures here.
+https://static.igem.org/mediawiki/parts/7/73/Hfq-serratia-frulhuq4.png
+Figure 1. Whole cell blots expressing OsmY-Hfq fusion protein under the control of the rhamnose promoter.
+https://static.igem.org/mediawiki/parts/8/81/Allsupernatant_blots2.png
+Figure 2. Supernatant blots of OsmY-HA (<partinfo>BBa_K1963007</partinfo>), OsmY-Hfq (E. coli- <partinfo>BBa_K1963008</partinfo>) and OsmY-Hfq (Serratia <partinfo>BBa_K1963009</partinfo>).. Western blotting with anti-HA antibody shows secretion of OsmY-HA, and OsmY-Hfq (E. coli) and anti-RNAP antibody shows no antigen detected in the supernatant therefore the cells remained intact and there was no cell lysis.
-https://static.igem.org/mediawiki/parts/4/49/Hfq-serratia-frulhuq3.png