Difference between revisions of "Part:BBa K314104"

Latest revision as of 20:05, 24 October 2010

T7 expression cassette

Lactose (IPTG) inducible T7 protein expression insert includes the f1 origin BBa_K314110, a Lac I generator BBa_K314111, a T7 promoter BBa_K314112 that includes the Elowitz standard RBS BBa_B0034. When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.

Usage and Biology

BBa_E0040 (GFP) was inserted between S and P using standard BioBrick assembly. The resulting constructs were grow in DH5a (constitutive promoters BBa_J23100 or BBa_J23114, as well as inducible promoter BBa_R0011) or BL21(DE3)* (T7 promoter, BBa_K314104) cells overnight at 298K in either the presence or absence of 0.5mM IPTG. The cells were harvested, washed in PBS, and then fluorescence for the whole cells was measured on a fluorometer (ex. 485, em. 525) and normalized to cell density (OD600). Cells that did not contain GFP were used as a blank. As observed, the addition of IPTG significantly increased the observed fluorescence of the cells.

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
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 126
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K314104 short</partinfo>
-Lac induced T7 protein expression insert includes f1 origin, Lac I gene and RBS
+Lactose (IPTG) inducible T7 protein expression insert includes  the f1 origin <partinfo>BBa_K314110</partinfo>, a Lac I generator <partinfo>BBa_K314111</partinfo>,  a T7 promoter <partinfo>BBa_K314112</partinfo> that includes the  Elowitz standard RBS <partinfo>BBa_B0034</partinfo>.  When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.
+===Usage and Biology===
+{| cellpadding="1" cellspacing="8"
+|-
+|[[Image:Washington2010_vector_assay_data.png | 650px]]
+|<partinfo>E0040</partinfo> (GFP) was inserted between S and P using standard BioBrick assembly.  The resulting constructs were grow in DH5a (constitutive promoters <partinfo>BBa_J23100</partinfo> or <partinfo>BBa_J23114</partinfo>, as well as inducible promoter <partinfo>R0011</partinfo>) or BL21(DE3)* (T7 promoter, <partinfo>BBa_K314104</partinfo>) cells overnight at 298K in either the presence or absence of 0.5mM IPTG.  The cells were harvested, washed in PBS, and then fluorescence for the whole cells was measured on a fluorometer (ex. 485, em. 525) and normalized to cell density (OD600).  Cells that did not contain GFP were used as a blank.  As observed, the addition of IPTG significantly increased the observed fluorescence of the cells.
+|-
+|}
 <!-- Add more about the biology of this part here