Difference between revisions of "Part:BBa K2933012"
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===Usage and Biology=== | ===Usage and Biology=== | ||
Flavobacterium johnsoniae (formerly Cytophaga johnsonae) is an environmental bacterium that can cause skin lesions in fish and is a plant pathogen. | Flavobacterium johnsoniae (formerly Cytophaga johnsonae) is an environmental bacterium that can cause skin lesions in fish and is a plant pathogen. | ||
+ | ==References== | ||
+ | |||
+ | |||
+ | ===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.<br> | ||
+ | <p style="text-align: center;"> | ||
+ | [[File:NDM-23-PCR.png|200px]]<br> | ||
+ | '''Figure 1.''' Left: The PCR result of JOHN-1. Right: 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 08:59, 8 September 2019
subclass B1 metallo-beta-lactamase JOHN-1, codon optimized in E. coli
This part encodes a protein called JOHN-1, which is a metallo-beta-lactamase of subclass B1.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 76
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 76
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 76
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 76
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 600
Usage and Biology
Flavobacterium johnsoniae (formerly Cytophaga johnsonae) is an environmental bacterium that can cause skin lesions in fish and is a plant pathogen.
References
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.
Figure 1. Left: The PCR result of JOHN-1. Right: 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.