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pcDNA4c/His-Xpress-HA-hBIG2(WT) (#RDB15364)

Expression vector of human BIG2, wild type.


Drawn by SnapGene® software
Sequence information
(Assembled from experimentally sequenced data)		 GenBank Flat File Format open
SnapGene file download
Publication Ishizaki, R., Mol. Biol. Cell 19 (6): 2650-2660 (2008). PMID 18417613. [PubMed] [Article] [RRC of NBRP]
Test sheet Data Sheet open 
 
Alternative name N7
Clone info. Expression vector of human Arf guanine nucleotide-exchange factors 2 (BIG2), wild type, tagged with Xpress and HA epitope at N-ter. This clone contains a cDNA fragment corresponding to the 180nt to 5972nt of NM_006420.3. Known differences (at least) : C to T substitution at 5342nt (Leu, synonym) ; A to G substitution at 5345nt (Lys, synonym).
Comment 1. BIG1 and BIG2 containing plasmids seem to be toxic to bacteria. After transformation, you need longer incubation time to get colonies as compared with usual transformed bacteria. Moreover, the colonies are smaller than usual ones. So, please take small colonies for liquid culture (The depositor is usually using DH5alpha for transformation).
2. Mini-prep: For liquid culture, you should use very fresh colonies otherwise you will get very low amounts of plasmids.
3. Maxi-prep (Large scale prep): Do not make a preculture from a single colony for large scale culture. You will get very low amounts of plasmids. The depositor would recommend you that after transformation and recovery culture, bring all transformed bacteria directly to large scale culture (200 ml or more of ampicillin containing medium). This helps a lot to get plasmids.
Expression was confirmed by the depositor with immunostaining.
Vector backbone pcDNA4/HisMax (plasmid)
Size of vector backbone 5.3 kb
Selectable markers Amp^r (E. coli), Zeo^r (mammalian cells).
Growth conditions LB+Amp, 37oC
Gene/insert name human BIG2 cDNA
Depositor|Developer Nakayama, Kazuhisa |

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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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 (Ishizaki, R. et al. Mol. Biol. Cell, 19 (6): 2650-2660, 2008). RECIPIENT agrees to use the BIOLOGICAL RESOURCE only for academic research in the non-profit organization.
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。
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 (Ishizaki, R. et al. Mol. Biol. Cell, 19 (6): 2650-2660, 2008)。本件リソースは非営利機関にのみ提供し、利用範囲は学術研究に制限する。

Catalog # Resource name Shipping form Fee
RDB15364 pcDNA4c/His-Xpress-HA-hBIG2(WT) DNA solution

How to cite this biological resource

Materials & Methods section:

The pcDNA4c/His-Xpress-HA-hBIG2(WT) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15364).

Reference section:

Ishizaki, R., Shin, H.W., Mitsuhashi, H., Nakayama, K., Redundant roles of BIG2 and BIG1, guanine-nucleotide exchange factors for ADP-ribosylation factors in membrane traffic between the trans-Golgi network and endosomes. Mol. Biol. Cell 19 (6): 2650-2660 (2008). PMID 18417613. [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 Ishizaki, R., Redundant roles of BIG2 and BIG1, guanine-nucleotide exchange factors for ADP-ribosylation factors in membrane traffic between the trans-Golgi network and endosomes. Mol. Biol. Cell 19 (6): 2650-2660 (2008). PMID 18417613. [PubMed] [Article] [RRC of NBRP]