Difference between revisions of "Part:BBa K3286100"

Latest revision as of 13:15, 21 October 2019

Expression construct for proteins in E. coli BL21DE3

This construct can be used to produce high amounts of protein in E. coli BL21DE3 (or another strain containing a T7 polymerase BBa_K1614000). Due to the Bicistronic Design (BCD) system BBa_M36516 [1], protein production is increased. Recombinant protein can be isolated using the Strep tag (BBa_K3286101), which can be removed with TEV (Tomato Etch Virus) protease at the TEV protease recognition site (BBa_K2927004). Integrate the protein of interest directly after the strep tag and make sure it is in frame. Also, remove its start codon.

[1] V. K. Mutalik et al., “Precise and reliable gene expression via standard transcription and translation initiation elements,” Nat. Methods, vol. 10, no. 4, pp. 354–360, Apr. 2013.

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 3: / Line 3: @@
 <partinfo>BBa_K3286100 short</partinfo>
-This construct can be used to produce high amounts of protein in E. coli BL21DE3 (or another strain containing a T7 polymerase). Due to the bicistronic design (BCD) system [1], protein production is increased. Recombinant protein can be isolated using the Strep tag, which can be removed with TEV protease at the TEV (Tomato Etch Virus) protease recognition site. Integrate the protein of interest directly after the strep tag and make sure it is in frame. Also, remove its start codon.
+This construct can be used to produce high amounts of protein in E. coli BL21DE3 (or another strain containing a T7 polymerase <partinfo>BBa_K1614000</partinfo>). Due to the Bicistronic Design (BCD) system <partinfo>BBa_M36516</partinfo> [1], protein production is increased. Recombinant protein can be isolated using the Strep tag (<partinfo>BBa_K3286101</partinfo>), which can be removed with TEV (Tomato Etch Virus) protease at the TEV protease recognition site (<partinfo>BBa_K2927004</partinfo>). Integrate the protein of interest directly after the strep tag and make sure it is in frame. Also, remove its start codon.
 [1] V. K. Mutalik et al., “Precise and reliable gene expression via standard transcription and translation initiation elements,” Nat. Methods, vol. 10, no. 4, pp. 354–360, Apr. 2013.