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pET28/EqtII(8-69)-EGFP-His6 (#RDB13497)

Marker of sphingomyelin in cell membrane. Bacterial expression plasmid of nontoxic Eqinatoxin II.


Drawn by SnapGene® software
Sequence information
(Assembled from experimentally sequenced data)		 GenBank Flat File Format open
SnapGene file download
Publication Makino, A., FASEB J. 29 (2): 477-493 (2015). PMID 25389132. [PubMed] [Article] [RRC of NBRP]
Test sheet Data Sheet open 
 
Clone info. Marker of sphingomyelin in cell membrane. Bacterial expression plasmid of nontoxic Eqinatoxin II. To use the recombinant protein, oxidization after the purification is required to prevent its cytolytic activity. Refer Abe, M., Methods Cell Biol. 137: 15-24 (2017).
Comment V8C K69C mutations. Optimized for expression in E. coli.
Please refer to the following note by the depositor:
- the expression of this protein was lesser than other protein expression vectors,
- induction was performed at 30 centigrade OVERNIGHT,
- the volume of cultivation was 500 to 1000 mL to obtain enough amount.
Please refer the Methods Cell Biol. 137 :15-24, 2017 (PMID: 28065303) for the method of expression and purification of the protein.
Vector backbone pET-28b (Plasmid)
Size of vector backbone 5.4 kb
Selectable markers Kan^r
Growth conditions LB + Kan, 37 oC
Gene/insert name Actinia equina Equinatoxin II cDNA Aequorea victoria GFP cDNA
Depositor|Developer Kobayashi, Toshihide | Abe, Mitsuhiro |

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
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MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ]
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Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested. (Makino, A., FASEB J. 29 (2): 477-493, 2015)
提供案内 (日本国内) [open/close]

必要書類
提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
手続きの概要は、「提供申込みについて[link]」をご覧ください。営利目的利用についてはお問い合わせください。
MTAに書く使用条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する(Makino, A., FASEB J. 29 (2): 477-493, 2015)。

Catalog # Resource name Shipping form Fee
RDB13497 pET28/EqtII(8-69)-EGFP-His6 DNA solution JPY 9,460 (not-for-profit academic purpose)
plus cost of shipping containers, dry ice (if required) and shipping charge

How to cite this biological resource

Materials & Methods section:

The pET28/EqtII(8-69)-EGFP-His6 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13497).

Reference section:

Makino, A., Abe, M., Murate, M., Inaba, T., Yilmaz, N., Hullin-Matsuda, F., Kishimoto, T., Schieber, N.L., Taguchi, T., Arai, H., Anderluh, G., Parton, R.G., Kobayashi, T., Visualization of the heterogeneous membrane distribution of sphingomyelin associated with cytokinesis, cell polarity, and sphingolipidosis. FASEB J. 29 (2): 477-493 (2015). PMID 25389132. [PubMed] [Article] [RRC of NBRP]

Further references such as user reports and related articles (go to bottom)

References

Original, user report and related articles

original Makino, A., Visualization of the heterogeneous membrane distribution of sphingomyelin associated with cytokinesis, cell polarity, and sphingolipidosis. FASEB J. 29 (2): 477-493 (2015). PMID 25389132. [PubMed] [Article] [RRC of NBRP]
protocol Abe, M., Dynamics of sphingomyelin-and cholesterol-enriched lipid domains during cytokinesis. Methods Cell Biol. 137: 15-24 (2017). PMID 28065303. [PubMed] [Article]
user_report Ueda, Y., Sphingomyelin localization in the intestinal crypt surface. Biochem Biophys Res Commun. 2022 Jun 30;611:14-18 (2022). PMID 35472605. [PubMed] [Article] [RRC of NBRP]