Difference between revisions of "Part:BBa K2933253"
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2933253 parameters</partinfo> | <partinfo>BBa_K2933253 parameters</partinfo> | ||
+ | <!-- --> | ||
+ | ===Usage and Biology=== | ||
+ | |||
+ | GIM-2 is a new variant of GIM-1 with a single mutation, A290G, which was recently discovered. | ||
+ | The German imipenemase-1 (GIM-1) MBL was first identified in clinical isolates of Pseudomonas aeruginosa in Germany in 2002. | ||
+ | Recently, GIM-1 has been identified in other bacterial species, such as Serratia marcescens, Enterobacter cloacae, and Acinetobacter pittii , indicating that it is transmitted on mobile genetic elements. | ||
+ | As a typical type of metallo-beta-lactamases which make bacteria antibiotic-resistant, it can hydrolyze extensive substrate and may pose a threat to human life and health. | ||
+ | |||
+ | |||
+ | <!-- --> | ||
+ | <span class='h3bb'>Sequence and Features</span> | ||
+ | <partinfo>BBa_K2933011 SequenceAndFeatures</partinfo> | ||
+ | |||
+ | |||
+ | |||
+ | ==Experimental results== | ||
+ | ===Molecular cloning=== | ||
+ | <p style="text-align: center;"> | ||
+ | [[File:GIM-2-PCR.jpeg|400px|]] [[File:GIM-2-veri.jpeg|250px|]]<br> | ||
+ | |||
+ | '''Figure 1.''' Left: The result of PCR, Right:The result of double enzyme digestion verification | ||
+ | |||
+ | |||
+ | |||
+ | ==References== | ||
+ | 1. Skagseth S , Akhter S , Paulsen M H , et al. Metallo-β-lactamase inhibitors by bioisosteric replacement: Preparation, activity and binding[J]. European Journal of Medicinal Chemistry, 2017, 135:159-173. | ||
+ | |||
+ | 2. Wendel AF, MacKenzie CR. 2015. Characterization of a novel metallo-lactamase variant, GIM-2, from a clinical isolate of Enterobacter cloacae in Germany. Antimicrob Agents Chemother 59:1824 –1825. | ||
+ | |||
+ | 3. Borra P S , Samuelsen O , Spencer J , et al. Crystal Structures of Pseudomonas aeruginosa GIM-1: Active-Site Plasticity in Metallo-beta-Lactamases[J]. Antimicrobial Agents and Chemotherapy, 2013, 57(2):848-854. | ||
+ | |||
+ | 4. Susann S, Trine J C, Gro Elin K B, James S, Ørjan S, Hanna-Kirsti S. L. Role of Residues W228 and Y233 in the Structure and Activity of Metallo-β-Lactamase GIM-1. Antimicrobial Agents and Chemotherapy Jan 2016, 60 (2) 990-1002 | ||
+ | |||
+ | |||
+ | |||
+ | <!-- Uncomment this to enable Functional Parameter display | ||
+ | ===Functional Parameters=== | ||
+ | <partinfo>BBa_K2933011 parameters</partinfo> | ||
<!-- --> | <!-- --> |
Revision as of 11:15, 21 September 2019
RBS b+Linker h+His+Linker a+Sumo+Linker b+GIM-2+T7 terminator
This part consists of RBS, protein coding sequence(His+Linker a+Sumo+Linker b+GIM-2) 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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 298
Illegal PstI site found at 1128 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 298
Illegal NheI site found at 75
Illegal NheI site found at 1158
Illegal PstI site found at 1128 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 298
Illegal BglII site found at 187
Illegal BamHI site found at 386 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 298
Illegal PstI site found at 1128 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 298
Illegal PstI site found at 1128 - 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
GIM-2 is a new variant of GIM-1 with a single mutation, A290G, which was recently discovered. The German imipenemase-1 (GIM-1) MBL was first identified in clinical isolates of Pseudomonas aeruginosa in Germany in 2002. Recently, GIM-1 has been identified in other bacterial species, such as Serratia marcescens, Enterobacter cloacae, and Acinetobacter pittii , indicating that it is transmitted on mobile genetic elements. As a typical type of metallo-beta-lactamases which make bacteria antibiotic-resistant, it can hydrolyze extensive substrate and may pose a threat to human life and health.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 731
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 731
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 731
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 731
- 1000COMPATIBLE WITH RFC[1000]
Experimental results
Molecular cloning
Figure 1. Left: The result of PCR, Right:The result of double enzyme digestion verification
References
1. Skagseth S , Akhter S , Paulsen M H , et al. Metallo-β-lactamase inhibitors by bioisosteric replacement: Preparation, activity and binding[J]. European Journal of Medicinal Chemistry, 2017, 135:159-173.
2. Wendel AF, MacKenzie CR. 2015. Characterization of a novel metallo-lactamase variant, GIM-2, from a clinical isolate of Enterobacter cloacae in Germany. Antimicrob Agents Chemother 59:1824 –1825.
3. Borra P S , Samuelsen O , Spencer J , et al. Crystal Structures of Pseudomonas aeruginosa GIM-1: Active-Site Plasticity in Metallo-beta-Lactamases[J]. Antimicrobial Agents and Chemotherapy, 2013, 57(2):848-854.
4. Susann S, Trine J C, Gro Elin K B, James S, Ørjan S, Hanna-Kirsti S. L. Role of Residues W228 and Y233 in the Structure and Activity of Metallo-β-Lactamase GIM-1. Antimicrobial Agents and Chemotherapy Jan 2016, 60 (2) 990-1002