Difference between revisions of "Part:BBa K3924017"
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==Profile== | ==Profile== | ||
Name: mchI<br/> | Name: mchI<br/> | ||
| − | Base Pairs: | + | Base Pairs: 210bp<br/> |
Origin: Escherichia coli, synthesis<br/> | Origin: Escherichia coli, synthesis<br/> | ||
Properties: Microcin H47(MccH47) immunity protein<br/> | Properties: Microcin H47(MccH47) immunity protein<br/> | ||
| + | |||
==Usage and Biology== | ==Usage and Biology== | ||
mchI was obtained from E. coli strain CA46. It is immune gene that encodes a immunity protein MchI against MccH47.<sup>[1]</sup> <br> | mchI was obtained from E. coli strain CA46. It is immune gene that encodes a immunity protein MchI against MccH47.<sup>[1]</sup> <br> | ||
==Design and Construction== | ==Design and Construction== | ||
We introduce mchI gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchI gene is added into lac operator(on RGP plasmid) so that the MchI can only be expressed induced by IPTG.Due to the time cost of obtaining the sequence, the sequence was submitted to the company for synthesis and we sequenced it to verify the correctness of the synthetic sequence. The sequence was added to RGP plasmid by Hifi DNA assembly replacing the TMAR sequence for further use.<br/> | We introduce mchI gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchI gene is added into lac operator(on RGP plasmid) so that the MchI can only be expressed induced by IPTG.Due to the time cost of obtaining the sequence, the sequence was submitted to the company for synthesis and we sequenced it to verify the correctness of the synthetic sequence. The sequence was added to RGP plasmid by Hifi DNA assembly replacing the TMAR sequence for further use.<br/> | ||
| − | [[Image:T--Tsinghua--part_mchI_construct.png|'''Figure 1: The design of mchI''']] | + | [[Image:T--Tsinghua--part_mchI_construct.png|center|600px|thumb|'''Figure 1: The design of mchI''']] |
Latest revision as of 07:13, 21 October 2021
mchI. It is immune gene that encodes a immunity protein MchI against MccH47
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 99
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 99
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 99
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 99
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 99
- 1000COMPATIBLE WITH RFC[1000]
Profile
Name: mchI
Base Pairs: 210bp
Origin: Escherichia coli, synthesis
Properties: Microcin H47(MccH47) immunity protein
Usage and Biology
mchI was obtained from E. coli strain CA46. It is immune gene that encodes a immunity protein MchI against MccH47.[1]
Design and Construction
We introduce mchI gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchI gene is added into lac operator(on RGP plasmid) so that the MchI can only be expressed induced by IPTG.Due to the time cost of obtaining the sequence, the sequence was submitted to the company for synthesis and we sequenced it to verify the correctness of the synthetic sequence. The sequence was added to RGP plasmid by Hifi DNA assembly replacing the TMAR sequence for further use.
Reference
[1] Vassiliadis G, Destoumieux-Garzón D, Lombard C, Rebuffat S, Peduzzi J. Isolation and characterization of two members of the siderophore-microcin family, microcins M and H47. Antimicrobial Agents and Chemotherapy. 2010 Jan;54(1):288-297. DOI: 10.1128/aac.00744-09. PMID: 19884380; PMCID: PMC2798501.