Please review the QC test results indicated by icon below as well as clone information before placing your order.

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

Expression vector of human BIG2, wild type.

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).
Gene/insert name	human BIG2 cDNA
Depositor|Developer	Nakayama, Kazuhisa |

 

Distribution information

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Ordering forms	Order form [Credit Card Payment] [Bank Transfer Payment] [Example of order form ] MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ] Please visit Information of Request for Distribution.[link]
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.

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

Please review the QC test results indicated by icon below as well as clone information before placing your order.

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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. [link to RRC of NBRP]

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

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QC test results

RIKEN BRC has sequenced portions of this material for quality test.
Please review the QC test results indicated by icon as well as clone information before placing your order.

Test sheet	RDB15364_A7I5p1-1.pdf

Nucleotide sequence of a portion of this resource (if available).

Primer: CMV_Forward (Pr0016) Region: CMV pro, QBI SP163, 6xHis, T7 tag, Xpress tag, HA, insert 5' Sequence file: RDB15364_A7I5a.seq
>D03588D1_2_A7I5_CMV-Forward_C06_09_ABI24.ab1     1 NNNNNNNNNN NNNTTGNNNC NNNGGGCGGT AGGCGTGTAC GGTGGGAGGT CTATATAAGC    61 AGAGCTCTCT GGCTAACTAG AGAACCCACT GCTTACTGGC TTATCGAAAT TAATACGACT   121 CACTATAGGG AGACCCAAGC TGGCTAGCGT TTAAACTTAA GCTTAGCGCA GAGGCTTGGG   181 GCAGCCGAGC GGCAGCCAGG CCCCGGCCCG GGCCTCGGTT CCAGAAGGGA GAGGAGCCCG   241 CCAAGGCGCG CAAGAGAGCG GGCTGCCTCG CAGTCCGAGC CGGAGAGGGA GCGCGAGCCG   301 CGCCGGCCCC GGACGGCCTC CGAAACCATG GGGGGTTCTC ATCATCATCA TCATCATGGT   361 ATGGCTAGCA TGACTGGTGG ACAGCAAATG GGTCGGGATC TGTACGACGA TGACGATAAG   421 GTACCTACCA TGTACCCGTA CGACGTGCCC GACTACGCGG GATCCACCAT GCAGGANAGC   481 CAGACCAAGA GCATGTTCGT GTCCCGGGCC CTGGANAAGA TCCTAGCCGA CAAGGANGTG   541 AAGCGGCCCC AGCACTCCCA GCTGCGCANG GCCTGCCAGG NGGCGCTCNA TGAAATTAAA   601 GCANAAATAG AAAANCANNN NNTNNGCACT GCTGCANNNN CNAANGNNAA ACTTCNTTGA 661 ANCTGANNNG NNTTTTCNNN NNTNNNANCT ANCNNGNNNN TNNNNNNNCN NNNNNNNNN //
Primer: BGH_rev2 (Pr0606) Region: BGH pA, insert 3' Sequence file: RDB15364_A7I5b.seq
>D03588D1_A7I5_2_BGH_rev2_D05_04_ABI08.ab1     1 NNNNNNNNNN NNNNNNNNNN NGGGGGNNGG GCANNNACNG ATGGCTGGCA ACTAGAAGGC    61 ACAGTCGAGG CTGATCAGCG GGTTTAAACG GGCCCTCTAG ACTCGAGCGG CCGCAAACGT   121 TTTTTTTTTT TTTTTTTATT TTAGGTGGCA GTTAGCCGTT ACAAGAAGTT TTATTAACAT   181 ATTCTCTAAT AATCACAAAC CTATACAGCT ATGTTCAGTA TTTTCAAGGA GAGTGACTTC   241 TTACAAAACG TTTATAGCCA ATCATGGGAT TTTGCTAGAA AACGATGGCA TATGCAATGA   301 CAGATTTGAC AGTTTAATCA NAAAAATGGA ATGATAGAAA TGAAGCANAC TCCCCACCTA   361 CTGTGCTAAC TGGGTATGCA ACAGTTTAAT GAAATACCAT AAGCCTTAGT TGGAGACTGT   421 GATGGAACAG CGTANAGGCC ATCCGTTTCC AGGCCATTCT GAGATCCAAG ATGCCAACTC   481 CTTGTTTCTA TGAAACTTCA CGAGATGTGC CAGTCAGAAA TGGCATGCTG AACATTCTGC   541 AGAGCTGCAG CACTGGCCTG GGCAGCCAGG GCTACCACAC TGGTGACAGT GCTGCTGGTA   601 CCTGTGATGG CTCTTCTGGT ATCCATATCT TATACACAAC ACCTATCCGT AGGAAGAACT   661 TCCGCANAAC TGCTCGGAGC TCANGGATCA GGTCAAACTG CATAATTTCA CACAAGTAGG   721 GGTAGTACAT TGAAGCATGT GCTTTGAACT TTTCATCATT TATCTTANNG GTTTTAGTTA   781 GAAGTAACNA CAAGAGANTT GTCCAGGCCT NCCGATGGCT CTCANAATTC ACAGNGNTGA   841 AATAGGNNAN ANCTTCNNTG CAAACNGTTN AAAGTCTCTG CTGNATTTCT NNCNNNGANN   901 CCNNGCNGNT CNNATNNNNA TACATTNNAA ANNGGNNNNT CAAANNNNNN NCNNNGNNNN 961 NNNNTNNNNN NTNNNNNNNN NGGNNNNNNN NTNNNNNNNN NN //
Primer: hBIG2_F1 (Pr0344) Region: insert mid (E738 WT) Sequence file: RDB15364_A7I5c.seq
>D03588D1_A7I5_2_hBIG2_F1_C02_03_ABI08.ab1     1 NNNNNNNNNN NNNNNNNNNT TCTCNNGGNN CAGGGCATGC TGGGAACGTC AGTTGAAGAC    61 ATAGCCCAAT TCCTGCACCA GGAGGAGCGC CTGGATTCCA CCCAAGTAGG CGATTTTCTG   121 GGAGATAGCG CAAGGTTCAA CAAGGAGGTG ATGTATGCCT ACGTGGACCA ACTTGACTTC   181 TGTGAAAAAG AATTTGTCTC AGCCCTGCGG ACATTCCTAG AAGGTTTCCG CCTACCTGGA   241 GAAGCCCAAA AGATTGACCG ATTAATGGAG AAGTTTGCCG CAAGATACAT AGAATGCAAC   301 CAAGGGCAAA CTCTGTTTGC TAGTGCTGAC ACTGCTTATG TCCTAGCGTA TTCAATTATT   361 ATGCTGACTA CAGACTTGCA CAGTCCTCAG GTAAAAAATA AAATGACGAA AGAGCAGTAT   421 ATTAAAATGA ATCGGGGTAT CAATGATAGT AAAGATCTGC CAGAAGAGTA TCTCTCAAGC   481 ATCTATGAAG AGATAGAAGG CAAGAAAATT GCAATGAAAG AAACAAAAGA GCTAACGATT   541 GCAACCAAAT CTACTAAGCA GAATGTAGCT AGTGAAAAGC AGCGGCGGCT GCTGTACAAC   601 TTAGAGATGG AGCAAATGGC TAAAACAGCC AAAGCTCTGA TGGAGGCTGT GAGCCATGCC   661 AAAGCCCCGT TTACCAGTGC CACTCACCTG GACCATGTCC GGCCAATGTT CNAACTGGTG   721 TGGACGCCAC TATTGGCAGC CTACAGCATC GGACTCCAGA ACTGTGATGA CACTGAAGTG   781 GCCTCCTTGT GTTTGGAANG CATCCGATGT GCAATCCGAN TCGCCTGCAT CTTTGGAATG   841 CAGCTGGAAC GANATGCCTA TGTTCANGCT CTTGCTCGCT TCTCCNNACT CNNNGCNNGC   901 TCNNGCATCN CAGAANTGNA NNANAAAAAC ATCNACNCCN TTAANANGCT TANCACANNG 961 GCNCANNCCG ANGGNNNNNA NCNNNNGNNN NNNNNNN //
Primer: hBIG2_R1 (Pr1019) Region: insert 5', HA, Xpress tag, T7 tag, 6xHis, QBI SP163 Sequence file: RDB15364_A7I5d.seq
>D03588D1_A7I5_2_hBIG2_R1_E02_05_ABI08.ab1     1 NNNNNNNNNN NNNNNNNGNA GCCTNGGCAG TGGTTTGNTT GATGAGANTT TTGCTGGCCA    61 AATAGATATT GTAACATGTT CTCACTGTCT GCAGGATAGT ACCCTCATGA ATTTCAATGT   121 GTGGGGAAGT CACTGCAGTC AGAAGAGCCT TAATTATTTG TAACTGAACC CCTTCATCAG   181 TCTGAGGGCC CTGAAAACAA CTGCAAATGG TTTCAACAAT TCTGTCGATC AGCCGCTTCC   241 CAGGGGCTCC ACTGTCAGGG GCGTTGCCAG TGATGTGCCC GTATGCGATG AGTTTCTGCA   301 AGCAGTCAAG GGATGTGCTG ACTACCCTTG GGGACTTGGA CTGGCAAGCT AGCTCGAATG   361 GAAGAAAATA CTTGTCAGCT TCAATGAAGT TTGCCTTTGG TGGTGCAGCA GTGCCAAGCC   421 TCTGCTTTTC TATTTCTGCT TTAATTTCAT CGAGCGCCAC CTGGCAGGCC CTGCGCAGCT   481 GGGAGTGCTG GGGCCGCTTC ACCTCCTTGT CGGCTAGGAT CTTCTCCAGG GCCCGGGACA   541 CGAACATGCT CTTGGTCTGG CTCTCCTGCA TGGTGGATCC CGCGTAGTCG GGCACGTCGT   601 ACGGGTACAT GGTAGGTACC TTATCGTCAT CGTCGTACAG ATCCCGACCC ATTTGCTGTC   661 CACCAGTCAT GCTAGCCATA CCATGATGAT GATGATGATG AGAACCCCCC ATGGTTTCGG   721 ANGCCGTCCG GGGCCGGCGC GGCTCGCGCT CCCTCTCCGG CTCGGACTGC GANGCAGCCC   781 GCTCTCTTGC GCGCCTTGNC GGGCTCCTCT CCCTTCTGGA ACCGAGGCCC GGGCCGGGGC   841 CTGNCTGCCG CTCGGCTGCC CNNAGCCTCT GCGCTAANCT TAAGTTTAAA CGCTAGCCAG   901 CTNNNNNNTN CCNATAGTGA GTNGNANTAA TTTCNATNAG CCAGTNANCA GNGGGNNCTC 961 NANTNANNNN NANNNCTCNG NNNANNNNNN NNNN //
Primer: hBIG2_F2 (Pr1020) Region: insert 3' Sequence file: RDB15364_A7I5e.seq
>D03588D1_A7I5_2_hBIG2_F2_F02_06_ABI08.ab1     1 NNNNNNNNNN NNNNNNNNNN NNNNNTCTTC NNNNNNAGAA ACCAGCAGCC TGGCCTGTTG    61 TTTGAGGNNC CTGTTTCGAA TGTATGTTGA TGAGAACCGC AGGGNTTCCT GGGAAGAAAT   121 ACAGCAGAGA CTTTTAACTG TTTGCAGTGA AGCTCTTGCC TATTTCATCA CTGTGAATTC   181 TGAGAGCCAT CGGGAGGCCT GGACAAGTCT CTTGTTGTTA CTTCTAACTA AAACCCTTAA   241 GATAAATGAT GAAAAGTTCA AAGCACATGC TTCAATGTAC TACCCCTACT TGTGTGAAAT   301 TATGCAGTTT GACCTGATCC CTGAGCTCCG AGCAGTTCTG CGGAAGTTCT TCCTACGGAT   361 AGGTGTTGTG TATAAGATAT GGATACCAGA AGAGCCATCA CAGGTACCAG CAGCACTGTC   421 ACCAGTGTGG TAGCCCTGGC TGCCCAGGCC AGTGCTGCAG CTCTGCAGAA TGTTCAGCAT   481 GCCATTTCTG ACTGGCACAT CTCGTGAAGT TTCATAGAAA CAAGGAGTTG GCATCTTGGA   541 TCTCAGAATG GCCTGGAAAC GGATGGCCTC TACGCTGTTC CATCACAGTC TCCAACTAAG   601 GCTTATGGTA TTTCATTAAA CTGTTGCATA CCCAGTTAGC ACAGTAGGTG GGGAGTCTGC   661 TTCATTTCTA TCATTCCATT TTTCTGATTA AACTGTCAAA TCTGTCATTG CATATGCCAT   721 CGTTTTCTAG CAAAATCCCA TGATTGGCTA TAAACGTTTT GTAAGAAGTC ACTCTCCTTG   781 AAAATACTGA ACATAGCTGT ATAGGTTTGT GANTATTAGA GAATATGTTA ATAAAACTTC   841 TTGTAACGGC TANNTGCNAC CNNANNNAAA AAAAAAAAAA AAAAACGTTT GCGGNNGCTC   901 GNNTCTANAG GNCCCGTTNN NNCCGCTNNN CANCNNCNNN NNNGNCTTNN ANTTGNNNNC 961 NNTCNNNNNN TNNNCNNNNN NNNNN //
Primer: SV40pro_F (Pr0110) Region: EM7 pro, ZeoR, SV40 pA Sequence file: RDB15364_A7I5f.seq
>D03588D1_A7I5_2_SV40pro_F_D02_04_ABI08.ab1     1 NNNNNNNNNN NNNNNNNNNN NGANNNGTGN NGNNGCTTTT TTGGANGCCT AGGCTTTTGC    61 AAAAAGCTCC CGGGAGCTTG TATATCCATT TTCGGATCTG ATCAGCACGT GTTGACAATT   121 AATCATCGGC ATAGTATATC GGCATAGTAT AATACGACAA GGTGAGGAAC TAAACCATGG   181 CCAAGTTGAC CAGTGCCGTT CCGGTGCTCA CCGCGCGCGA CGTCGCCGGA GCGGTCGAGT   241 TCTGGACCGA CCGGCTCGGG TTCTCCCGGG ACTTCGTGGA GGACGACTTC GCCGGTGTGG   301 TCCGGGACGA CGTGACCCTG TTCATCAGCG CGGTCCAGGA CCAGGTGGTG CCGGACAACA   361 CCCTGGCCTG GGTGTGGGTG CGCGGCCTGG ACGAGCTGTA CGCCGAGTGG TCGGAGGTCG   421 TGTCCACGAA CTTCCGGGAC GCCTCCGGGC CGGCCATGAC CGAGATCGGC GAGCAGCCGT   481 GGGGGCGGGA GTTCGCCCTG CGCGACCCGG CCGGCAACTG CGTGCACTTC GTGGCCGAGG   541 AGCAGGACTG ACACGTGCTA CGAGATTTCG ATTCCACCGC CGCCTTCTAT GAAAGGTTGG   601 GCTTCGGAAT CGTTTTCCGG GACGCCGGCT GGATGATCCT CCAGCGCGGG GATCTCATGC   661 TGGAGTTCTT CGCCCACCCC AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA   721 ATAGCATCAC AAATTTCACA AATAAAGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT   781 CNNAACTCAT CAATGTATCT TATCATGTCT GTATACCGTC GACCTCTANC TAGAGCTTGG   841 CGTANTCATG GTCATAGCTG TTTCCTGTGT GAAATTGTTA TCCGCTCANN NTTCCNCACA   901 ACATACNANC NNNAAGCNTA AAGNGTAAAG CCNGGGGGNN NCNNATGAGT GANCTAACTC 961 ACNTTNANNG CGNNGNNNNN NCNNNCNNCT TTCCANNNNG NNANNNNNNN NNN //

Please visit Sequencing and PCR primers for primer information.

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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. [link to RRC of NBRP]

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• human BIG2

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External Database
human BIG2

• in KEGG Gene
• in KEGG Orthology
• in NCBI Gene

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