Difference between revisions of "Part:BBa K2933178"
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<partinfo>BBa_K2933178 parameters</partinfo> | <partinfo>BBa_K2933178 parameters</partinfo> | ||
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+ | ===Usage and Biology=== | ||
+ | <br> | ||
+ | ===Molecular cloning=== | ||
+ | |||
+ | First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.We then extracted plasmids and performed double enzyme digestion verification.<br> | ||
+ | <p style="text-align: center;"> | ||
+ | [[File:BlaB-14-PCR.png|300px]]<br> | ||
+ | '''Figure 1.''' (a) The PCR result of BlaB-14. (b) The verification results by enzyme digestion.<br> | ||
+ | </p> | ||
+ | After verification, it was determined that the construction is successful. We converted the plasmid to ''E. coli'' BL21(DE3) for expression and purification. <br> |
Revision as of 12:51, 21 September 2019
Tac promoter+RBS a+Linker g+GST+Linker e+BlaB-14
This part consists of Tac promoter,RBS and protein coding sequence (GST+Linker e+BlaB-14),and the biological module can be built into E.coli for protein expression.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1355
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 181
Usage and Biology
Molecular cloning
First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.We then extracted plasmids and performed double enzyme digestion verification.
Figure 1. (a) The PCR result of BlaB-14. (b) The verification results by enzyme digestion.
After verification, it was determined that the construction is successful. We converted the plasmid to E. coli BL21(DE3) for expression and purification.