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]

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 <partinfo>BBa_K314104 short</partinfo>
-Lac induced protein expression insert includes f1 origin K314110, Lac I generator K314111, a T7 promoter k314112, and the Elowitz standard RBS B0034.  When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.
+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.
-Lac induced T7 promoter, <partinfo>BBa_K314112</partinfo>, protein expression insert includes f1 origin [https://parts.igem.org/Part:BBa_K314110 K314110], a Lac I generator [https://parts.igem.org/Part:BBa_K314111 K314111],  a high expression promoter [https://parts.igem.org/Part:BBa_J23100 J23100], and the  Elowitz standard RBS [https://parts.igem.org/Part:BBa_B0034 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===