Difference between revisions of "Part:BBa K2933267"
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First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.<br> | First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.<br> | ||
<p style="text-align: center;"> | <p style="text-align: center;"> | ||
− | [[File: | + | [[File:TJUSLS China--Elbla2-1-PCR.png|600px]]<br> |
'''Figure 1.''' The PCR result of ElblaII. | '''Figure 1.''' The PCR result of ElblaII. | ||
==References== | ==References== | ||
[1] Girlich D, Poirel L, Nordmann P, Diversity of naturally occurring Ambler class B metallo-β-lactamases in Erythrobacter spp. The Journal of Antimicrobial Chemotherapy [31 Jul 2012, 67(11):2661-2664] | [1] Girlich D, Poirel L, Nordmann P, Diversity of naturally occurring Ambler class B metallo-β-lactamases in Erythrobacter spp. The Journal of Antimicrobial Chemotherapy [31 Jul 2012, 67(11):2661-2664] |
Latest revision as of 15:02, 28 September 2019
RBS+Linker h+His+Linker f+ElBlaII+T7 terminator
This part consists of RBS, Linker h, protein coding sequence(His+Linker f+ElBlaII) and T7 terminator,and the biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Unknown
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
This composite part is made up with six basic parts(RBS , Linker h, His, Linker f , ElBlaII and T7 terminator ). It encodes a protein which is ElblaII fused with His tag. The fusion protein is about 27.5 kD. It is convenient for us to purify our target protein.
Molecular cloning
First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.
Figure 1. The PCR result of ElblaII.