MspA-89M plasmid from Shuo Huang
MspA-89M
Cat. No.:
MC_0101206
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-89M (I89M/D90N/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 89. It only has a single amino acid difference at the site 89 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 89 is located within the pore constriction and faces to the pore’s exterior. The MspA-89M mutant is designed to map the potential reactive sites within MspA to form a nanoreactor. The structure of the mutant, and its sensing performance to a model reactant (tetrachloroaurate(III) ion) are investigated. It is revealed that site 89 is unavailable as a reactive site because it faces to pore’s exterior (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 | High Copy |
| 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.
Customers who viewed this item also viewed:
CRISPR
CRISPR
Constitutive expression of Cas9 together with inducible expression of lambda RED and sgR
CRISPR/Cas9-assisted gRNA-free one-step genome editing