Difference between revisions of "Part:BBa K525222"

Revision as of 21:07, 21 September 2011

S-layer cspB from Corynebacterium halotolerans

S-layer Corynebacterium halotolerans

Important parameters

Experiment	Characteristic	Result
Expression (E. coli)	Localisation	Cell membrane
	Compatibility	E. coli KRX
	Induction of expression	L-rhamnose for induction of T7 polymerase
	Specific growth rate (un-/induced)	0.245 h^-1 / 0.109 h^-1
	Doubling time (un-/induced)	2.83 h / 6.33 h

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1103
Illegal XhoI site found at 559
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 226
Illegal NgoMIV site found at 1315
Illegal AgeI site found at 217
Illegal AgeI site found at 458
Illegal AgeI site found at 505
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 907
Illegal BsaI.rc site found at 214
Illegal BsaI.rc site found at 592
Illegal BsaI.rc site found at 994

Expression in E. coli

The CspB gen was fused with a monomeric RFP (BBa_E1010) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization.

The CspB|mRFP fusion protein was overexpressed in E. coli KRX after induction of T7 polymerase by supplementation of 0,1 % L-rhamnose using the [http://2011.igem.org/Team:Bielefeld-Germany/Protocols/Downstream-processing#Expression_of_S-layer_genes_in_E._coli autinduction protocol] from promega.

Figure 1: Growthcurve of E. coli KRX expressing the fusion protein of CspB and mRFP with and without induction. A curve depicting KRX wildtype is shown for comparsion.

Figure 2: RFU to OD₆₀₀ ratio of E. coli KRX expressing the fusion protein of CspB and mRFP with and without induction. A curve depicting KRX wildtype is shown for comparsion.

@@ Line 12: / Line 12: @@
 |rowspan="5"|[[Part:BBa_K525222#Expression_in_E._coli | Expression (''E. coli'')]]
 |Localisation
-|Inclusion body
+|Cell membrane
 |-
 |Compatibility
-|''E. coli'' KRX and BL21(DE3)
+|''E. coli'' KRX
 |-
 |Induction of expression
-|Expression of T7 polymerase + IPTG, lactose
+|L-rhamnose for induction of T7 polymerase
 |-
 |Specific growth rate (un-/induced)
@@ Line 41: / Line 41: @@
 <partinfo>BBa_K525222 parameters</partinfo>
 <!-- -->
 ===Expression in ''E. coli''===
 The CspB gen was fused with a monomeric RFP ([https://parts.igem.org/Part:BBa_E1010 BBa_E1010]) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization.