mPiezo2-CMV-Sport6 | Addgene 81073 product information

This webpage contains legacy information. The product is either no longer available from the supplier or has been delisted at Labome.

product summary

company name :

Addgene

product type :

cDNA

product name :

mPiezo2-CMV-Sport6

catalog :

81073

citations: 3

Reference
Duan M, Jia Y, Huo L, Gao Y, Wang J, Zhang W, et al. Potentiation of PIEZO2 mechanically-activated currents in sensory neurons mediates vincristine-induced mechanical hypersensitivity. Acta Pharm Sin B. 2023;13:3365-3381 pubmed publisher
Huo L, Gao Y, Zhang D, Wang S, Han Y, Men H, et al. Piezo2 channel in nodose ganglia neurons is essential in controlling hypertension in a pathway regulated directly by Nedd4-2. Pharmacol Res. 2021;164:105391 pubmed publisher
Coste B, Murthy S, Mathur J, Schmidt M, Mechioukhi Y, Delmas P, et al. Piezo1 ion channel pore properties are dictated by C-terminal region. Nat Commun. 2015;6:7223 pubmed publisher

product information

Catalog Number :

81073

Product Name :

mPiezo2-CMV-Sport6

article :

doi	10.1038/ncomms8223
id	21963
pubmed_id	26008989

bacterial resistance :

Ampicillin

cloning :

backbone

pCMV-Sport6

backbone_mutation

backbone_origin

backbone_size

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Mammalian Expression

growth notes :

Please note- when compared to the Piezo2 reference sequence NM_001039485.4, the insert in this plasmid contains mutation D1896E and deletion of residues V1897 and E1898. Piezo2 was amplified out of mouse DRGs. The variant that is present in DRGs when it was cloned is missing 6 base pairs. Here is the link to the original manuscript the reported protein sequence is in the supplementary data. The plasmid functions as described. http://science.sciencemag.org/content/330/6000/55.full The depositor recommends the following protocol for researchers having issues growing or isolating plasmid 81073 (mPiezo2-CMV-Sport6): For Piezo2- inoculate culture from glycerol stock into 4-5mls of LB media. Grow the culture at 37 deg for 4-5 hrs and then transfer the culture to larger volumes of media (about 100-150mls) and grow overnight at 37 deg. Usual maxi prep yields are low in the range of 50-100ng/ul. We digest verify the preps by digesting with KpnI and NotI to give 8.2kb insert and 4.4kb vector bands or with KpnI and SpeI to give 2 bands with sizes 8.6 kb and 4.2 kb. When handling glycerol stocks for Piezo2, always re-streak and pick single clones. Never inoculate the glycerol stock into an overnight culture. Our lab has had issues with clones that grow with amp resistance but after digestion, show a prominent ~4 kb single band. After mini-prep or maxi-prep, always confirm the DNA product by restriction digest. We digest with KpnI and SpeI and this leads to 2 bands with sizes 8.6 kb and 4.2 kb. We digest verify every time we maxi prep. For our maxi preps, we notice that our yields are low - sometimes ~ 50-100 ng/ul when eluting in 1 ml from ~150 ml of culture. Alternatively, transform your DNA into XL-10 gold cells. Plate and grow for 3-4 days at 25C. Select single colonies and inoculate 200-300 mL of media. Grow with shaking at 25C for 3-4 days. Maxiprep the culture as normal, digest DNA to verify as above.

growth strain :

mouse Piezo2

growth temp :

transform your DNA into XL-10 gold cells. Plate and grow for 3-4 days at 25C. Select single colonies and inoculate 200-300 mL of media. Grow with shaking at 25C for 3-4 days.

origin :

pi :

alt_names

cloning

clone_method	Restriction Enzyme
cloning_site_3	NotI
cloning_site_5	KpnI
promoter
sequencing_primer_3
sequencing_primer_5	CGC AAA TGG GCG GTA GGC GTG
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	5930434P17, 9030411M15Rik, 9430028L06Rik, Fam38b, Fam38b2
gene	Piezo2
id	667742

genbank_ids

mutation

name

Piezo2 (please see depositor comments below)

shRNA_sequence

size

8475

species

10090

Mus musculus