MspA-90M plasmid from Shuo Huang
MspA-90M
Cat. No.:
MC_0101201
Availability:
4 µg of lyophilized plasmid
(1 µg for low-copy plasmid)
(1 µg for low-copy plasmid)
Price:
$65
| Keywords | Expression |
| Vector Backbone | pET-30a (+) |
| Source | Member Deposit |
| Depositor | Shuo Huang |
| Organization | Nanjing University |
| Description | The plasmid MspA-90M (D90M/D91N/D93N/D118R/D134R/E139K) is designed for the preparation of a mutant Mycobacterium smegmatis porin A (MspA) nanopore, which is engineered with a methionine at the site 90. It only has a single amino acid difference at the site 90 from the previously reported M2 MspA (D90N/D91N/D93N/D118R/D134R/E139K) (Proc. Natl. Acad. Sci., 2008, 105, 20647-20652), which has been applied in DNA sequencing (Nat. Biotechnol., 2012, 30, 349-353). MspA (PDB ID: 1UUN) possesses a narrow and short pore constriction, measuring ~1.2 nm at its narrowest spot and ~1.0 nm in an overall length. With this advantageous geometry, MspA exhibits an excellent sensing performance as a nanoreactor to probe single-molecule reactions (Nat. Commun., 2019, 10, 5668.; Chem. Sci., 2020,11, 879-887.; Anal. Chem. 2021, 93, 3, 1529–1536). The reported engineering of MspA to form corresponding nanoreactor has focused only on site 91. In our work, other potential engineering sites of MspA have been thoroughly investigated. The amino acid at the site 90 is located within the pore constriction and has a lumen-facing orientation. The MspA-90M mutant is designed to map the potential reactive sites within MspA to form a nanoreactor. The protein yield for this mutant is relatively low. The structure of the mutant, and its sensing performance to a model reactant (tetrachloroaurate(III) ion) are investigated. It is revealed that the sensing performance is inversely correlated to the cross-sectional diameter of the pore constriction size of the engineering site (ACS Sens. 2021. DOI: 10.1021/acssensors.1c00792.). |
| Publication | Zhang, J., Cao, J., Jia, W. et al. Mapping potential engineering sites of Mycobacterium smegmatis porin A (MspA) to form a nanoreactor. ACS Sensors 2021. DOI: 10.1021/acssensors.1c00792. |
| Plasmid Copy | Unknown |
| Bacterial Resistance | Kanamycin |
| Growth Strain | DH5-Alpha |
| Growth Temperature | 37°C |
| Plasmid Size (bp) | 5814 |
*This material may be covered by one or more patents, trademarks and/or copy rights owned or controlled by Depositors or any third parties. This material is available to academic and nonprofit organizations for research use only. Please contact MolecularCloud at plasmid@genscript.com and the Depositor directly if you are from industrial institute or attempt for profit application. This material is not intended to be used therapeutic or diagnostic purposed in humans or animals.
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