Difference between revisions of "Part:BBa K4241013"

(Overview and Usage and BIology)
Line 8: Line 8:
  
 
===Usage and Biology===
 
===Usage and Biology===
−
In order ot increase the yield of recombinant protein during production, the T7 Tag can be used. This part also contains thrombin and 6xHis for double His-pulldown purification.
+
In order to increase the yield of recombinant protein during production, the T7 Tag can be used. This part also contains thrombin and 6xHis for double His-pulldown purification.
 
It is of note that this seqence does not contain a start codon, therefore to use this part it must be in frame with a promoter with a Met initiation site.
 
It is of note that this seqence does not contain a start codon, therefore to use this part it must be in frame with a promoter with a Met initiation site.
 
<br/>
 
<br/>

Revision as of 10:16, 9 October 2022


6xHis-thrombin-T7Tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 55
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Overview

This contains adds the 6xHis-Thrombin-T7 tag. The T7 tag is proven to increase the yield of recombinant protein production, whilst thrombin is used to cleave the 6xHis tag for purification purposes.

Usage and Biology

In order to increase the yield of recombinant protein during production, the T7 Tag can be used. This part also contains thrombin and 6xHis for double His-pulldown purification. It is of note that this seqence does not contain a start codon, therefore to use this part it must be in frame with a promoter with a Met initiation site.

For purification:
1. His-pulldown complete construct
2. Cleave thrombin
3. His-pulldown once again to remove uncleaved product.