Difference between revisions of "Part:BBa K302012:Experience"

Revision as of 16:44, 26 October 2010

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Applications of BBa_K302012

Characterisation by Team Newcastle 2010

We integrated our part into the Bacillus subtilis 168 chromosome at amyE (using the integration vector pGFP-rrnB) and selected for integration by testing for the ability to hydrolyse starch. Homologous recombination at amyE destroys endogenous expression of amylase. Colonies that are not able to break down starch on agar plate do not have a white halo when exposed to iodine.

The part was co-transcribed with gfp fluorescent marker by transcriptional fusion after the yneA coding sequence.

We characterised the part first without, and then with, LacI repression (using the integration vector pMutin4 to integrate lacI into the Bacillus subtilis 168 chromosome).

Graph1:

Graph 1 shows that overexpression (no Lac repression) of the yneA gene (ΔamyE:pSpac(hy)-oid::yneA) leads to a longer cell length compared with our control Bacillus subtilis 168.

See below: Bacillus subtilis 168 cells (left),Bacillus subtilis expressing yneA(centre) and Bacillus subtilis overexpressing yneA(right)

Graph2:

Graph 2 shows the percentage of cells at different lengths(μm)uninduced

See below: Bacillus subtilis 168 cells (left) and non-induced cells(right)

Graph3:

Graph 3 shows the percentage of cells at different lengths(μm)induced at 0.2mM IPTG

See below: Bacillus subtilis 168 cells (left) and cells induced at 0.2mM IPTG(right)

Graph4:

Graph 4 shows the percentage of cells at different lengths(μm)induced at 1mM IPTG

See below: Bacillus subtilis 168 cells (left) and cells induced at 1mM IPTG(right)

Graphs 2,3 and 4 show a greater proportion of cells at a higher concentration of IPTG(1mM IPTG), compared with Bacillus subtilis 168 our control population.

User Reviews

UNIQ599cf2f876fc3108-partinfo-00000000-QINU UNIQ599cf2f876fc3108-partinfo-00000001-QINU

@@ Line 5: / Line 5: @@
 ===Applications of BBa_K302012===
-The filamentous cell part is repressed by LacI. The LacI repression is removed by induction with IPTG. Without LacI repression a higher proportion of the cells are filamentous. The filamentous cell phenotype is achieved by overexpressing ''yneA'' which is an indirect inhibitor of cell division.
+====Characterisation by Team Newcastle 2010====
+We integrated our part into the ''Bacillus subtilis'' 168 chromosome at ''amyE'' (using the integration vector pGFP-rrnB) and selected for integration by testing for the ability to hydrolyse starch. Homologous recombination at ''amyE'' destroys endogenous expression of amylase. Colonies that are not able to break down starch on agar plate do not have a white halo when exposed to iodine.
+The part was co-transcribed with ''gfp'' fluorescent marker by transcriptional fusion after the ''yneA'' coding sequence.
+We characterised the part first without, and then with, LacI repression (using the integration vector pMutin4 to integrate ''lacI'' into the ''Bacillus subtilis'' 168 chromosome).
-We characterised the part first without, and then with, LacI repression (using the integration vector pMutin4 to integrate ''lacI'' into the ''Bacillus subtilis'' 168 chromosome.
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