Resource data sheet
Prev.
Please review the QC test results indicated by check icon below as well as clone information before placing your order.

pcDNA4c/His-Xpress-HA-hBIG2(E738K)

Expression vector of human BIG2, dominant-negative mutant E738K.

Catalog number RDB15365
Resource name pcDNA4c/His-Xpress-HA-hBIG2(E738K)
Alternative name N8
Clone info. Expression vector of human Arf guanine nucleotide-exchange factors 2 (BIG2), dominant-negative mutant E738K, 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) : A to G substitution at 2390nt (Gly, synonym) ; A to G substitution at 2399nt (Gln, synonym) ; T to C substitution at 2405nt (Ile, synonym) ; C to T substitution at 2408nt (Asp, synonym) ; 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 |
Other clones in our bank

External Database
human BIG2

          Reference sequence
            
           

          Distribution information

          Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
          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.
          Ordering Please visit Information of Request for Distribution.[link] 
          Order form 
          [Credit Card Payment]  [Bank Transfer Payment]
          Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
          提供案内 (日本国内) [open/close]

          提供条件 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。
          提供依頼 手続きの詳細は、「提供申込みについて[link]」をご覧ください。
          提供依頼書 [Word]
          提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]

          Catalog # Resource name Shipping form Fee (non-profit org.)
          RDB15365 pcDNA4c/His-Xpress-HA-hBIG2(E738K) DNA solution

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

          How to cite this biological resource

          Materials & Methods section:

          The pcDNA4c/His-Xpress-HA-hBIG2(E738K) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15365).

          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.

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


          References and tips

          Featured content


          QC test results

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

          Test sheet RDB15365_A7I5p1-1.pdf check

          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: RDB15365_A7I5a.seq check
          >D03588D2_A7I5_2_CMV_Forward_A09_03_ABI24.ab1
              1 NNNNNNNNNN NNNNNTNNNN GCNNNNGGGC GGTANGCGTG TACGGTGGGN NGTCTATATA
             61 AGCAGAGCTC TCTGGCTAAC TAGAGAACCC ACTGCTTACT GGCTTATCGA AATTAATACG
            121 ACTCACTATA GGGAGACCCA AGCTGGCTAG CGTTTAAACT TAAGCTTAGC GCAGAGGCTT
            181 GGGGCAGCCG AGCGGCAGCC AGGCCCCGGC CCGGGCCTCG GTTCCAGAAG GGAGAGGAGC
            241 CCGCCAAGGC GCGCAAGAGA GCGGGCTGCC TCGCAGTCCG AGCCGGAGAG GGAGCGCGAG
            301 CCGCGCCGGC CCCGGACGGC CTCCGAAACC ATGGGGGGTT CTCATCATCA TCATCATCAT
            361 GGTATGGCTA GCATGACTGG TGGACAGCAA ATGGGTCGGG ATCTGTACGA CGATGACGAT
            421 AAGGTACCTA CCATGTACCC GTACGACGTG CCCGACTACG CGGGATCCAC CATGCAGGAG
            481 AGCCAGACCA AGAGCATGTT CGTGTCCCGG GCCCTGGAGA AGATCCTAGC CGACAAGGAG
            541 GTGAAGCGGC CCCAGCACTC CCAGCTGCGC AGGGCCTGCC AGGTGGCGCT CGATGAAATT
            601 AAAGCAGAAA TAGAAAAGCA GAGGCTTGGC ACTGCTGCAC CACCAAAGGC AAACTTCATT
            661 GAAGCTGACA AGTATTTTCT TCCNTTCGAG CTAGCTTGCC AGTCCAAGTC CCCAAGGGTA
            721 GTCAGCACAT CCCTTGACTG CTTGCAGAAA CTCATCGCAT ACGGGCACAT CACTGGCAAC
            781 GCCCCTGACA GTGGANCCCC NGGGAAGCGG CTGANCGACA GAATTGTTGA AACCATTTGC
            841 AGTTGTTTTC NGGNCCCTCN NANNGATGAA NGGGTTCAGT TACNAATNAT TNANNNNCNN
            901 CNGNNNGCNN NGNNNTCCCN NCNCNNNNNA ANTNNNNNNG NNNNNATCNN NNNNANNNNN
          961 NNNNN
          //
          Primer: BGH_rev2 (Pr0606)
          Region: BGH pA, insert 3'
          Sequence file: RDB15365_A7I5b.seq check
          >D03588D2_A7I5_2_BGH_rev2_F05_06_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNGNGNNN NNNCAANAAC AGNNGGCTGG CAACTAGAAG
             61 GCACAGTCGA GGCTGATCAG CGGGTTTAAA CGGGCCCTCT AGACTCGAGC GGCCGCAAAC
            121 GTTTTTTTTT TTTTTTTTTA TTTTAGGTGG CAGTTAGCCG TTACAAGAAG TTTTATTAAC
            181 ATATTCTCTA ATAATCACAA ACCTATACAG CTATGTTCAG TATTTTCAAG GAGAGTGACT
            241 TCTTACAAAA CGTTTATAGC CAATCATGGG ATTTTGCTAG AAAACGATGG CATATGCAAT
            301 GACAGATTTG ACAGTTTAAT CAGAAAAATG GAATGATAGA AATGAAGCAG ACTCCCCACC
            361 TACTGTGCTA ACTGGGTATG CAACAGTTTA ATGAAATACC ATAAGCCTTA GTTGGAGACT
            421 GTGATGGAAC AGCGTANAGG CCATCCGTTT CCAGGCCATT CTGAGATCCA AGATGCCAAC
            481 TCCTTGTTTC TATGAAACTT CACGAGATGT GCCAGTCAGA AATGGCATGC TGAACATTCT
            541 GCAGAGCTGC AGCACTGGCC TNNNNNNNCA GGGCTACCAC ACTGGTGACA GTGCTGCTGG
            601 TACCTGTGAT GGNTCTTCTG GTATCCATAT CTTATACNCA ACACCTATCC GTANNAAGAA
            661 CTTCCGCANA ACTGCTCNNA NCTCNNGNAT CNNNTCNAAC TGCANNNTTT CNCACANGTN
            721 NGGGNANNAN NTTGNANCNT GNGCTTTGNA CTTTTCNTCA TTTNNCNTNN NNNTTTANNT
            781 NNNANTNNCN NCNNNANANN NGTNCNNNNN TNCNNANGNN NNTCNNANNN NCNNNGNTNN
          841 ANTNNNNNNN NNN
          //
          Primer: hBIG2_F1 (Pr0344)
          Region: insert mid (E738K)
          Sequence file: RDB15365_A7I5c.seq check
          >D03588D2_A7I5_2_hBIG2_F1_C09_09_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNTNC TCNNGGNNCA NGGCATGCTG GGAACGTCAG TTGAAGANAT
             61 AGCCCAATTC CTGCACCAGG AGGAGCGCCT GGATTCCACC CAAGTAGGCG ATTTTCTGGG
            121 AGATAGCGCA AGGTTCAACA AGGAGGTGAT GTATGCCTAC GTGGACCAAC TTGACTTCTG
            181 TGAAAAAGAA TTTGTCTCAG CCCTGCGGAC ATTCCTAGAA GGTTTCCGCC TACCTGGGAA
            241 AGCCCAGAAG ATCGATCGAT TAATGGAGAA GTTTGCCGCA AGATACATAG AATGCAACCA
            301 AGGGCAAACT CTGTTTGCTA GTGCTGACAC TGCTTATGTC CTAGCGTATT CAATTATTAT
            361 GCTGACTACA GACTTGCACA GTCCTCAGGT AAAAAATAAA ATGACGAAAG AGCAGTATAT
            421 TAAAATGAAT CGGGGTATCA ATGATAGTAA AGATCTGCCA GAAGAGTATC TCTCAAGCAT
            481 CTATGAAGAG ATAGAAGGCA AGAAAATTGC AATGAAAGAA ACAAAAGAGC TAACGATTGC
            541 AACCAAATCT ACTAAGCAGA ATGTAGCTAG TGAAAAGCAG CGGCGGCTGC TGTACAACTT
            601 AGAGATGGAG CAAATGGCTA AAACAGCCAA AGCTCTGATG GAGGCTGTGA GCCATGCCAA
            661 AGCCCCGTTT ACCAGTGCCA CTCACCTGGA CCATGTCCGG CCAATGTTCA AACTGGTGTG
            721 GACGCCACTA TTGGCAGCCT ACAGCATCGG ANTCCAGAAC TGTGANGANA CTGAANTGGC
            781 CTCCTTGTGT TTGGAANGNA TCCGATGTGC AATCCGAATC GCNTGCATCT TTGGAATGCA
            841 GCTNGAACGA NNNGNCNATG TTCNNNNNCT NGNTCGNTTC NNCNNANNNN CANCNNGNNN
          901 CNGCNTCNCA NAAATGNANN CNNAAAAANN TCNNNNNNNN NN
          //
          Primer: hBIG2_R1 (Pr1019)
          Region: insert 5', HA, Xpress tag, T7 tag, 6xHis, QBI SP163
          Sequence file: RDB15365_A7I5d.seq check
          >D03588D2_A7I5_2_hBIG2_R1_H02_08_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN AGCCTTGGCA GTGGTTTGAT TGATGAGNNT TTTGCTGGCC
             61 AAATAGATAT TGTAACATGT TCTCACTGTC TGCAGGATAG TACCCTCATG AATTTCAATG
            121 TGTGGGGAAG TCACTGCAGT CAGAAGAGCC TTAATTATTT GTAACTGAAC CCCTTCATCA
            181 GTCTGAGGGC CCTGAAAACA ACTGCAAATG GTTTCAACAA TTCTGTCGAT CAGCCGCTTC
            241 CCAGGGGCTC CACTGTCAGG GGCGTTGCCA GTGATGTGCC CGTATGCGAT GAGTTTCTGC
            301 AAGCAGTCAA GGGATGTGCT GACTACCCTT GGGGACTTGG ACTGGCAAGC TAGCTCGAAT
            361 GGAAGAAAAT ACTTGTCAGC TTCAATGAAG TTTGCCTTTG GTGGTGCAGC AGTGCCAAGC
            421 CTCTGCTTTT CTATTTCTGC TTTAATTTCA TCGAGCGCCA CCTGGCAGGC CCTGCGCAGC
            481 TGGGAGTGCT GGGGCCGCTT CACCTCCTTG TCGGCTAGGA TCTTCTCCAG GGCCCGGGAC
            541 ACGAACATGC TCTTGGTCTG GCTCTCCTGC ATGGTGGATC CCGCGTAGTC GGGCACGTCG
            601 TACGGGTACA TGGTAGGTAC CTTATCGTCA TCGTCGTACA GATCCCGACC CATTTGCTGT
            661 CCACCAGTCA TGCTAGCCAT ACCATGATGA TGATGATGAT GAGAACCCCC CATGGTTTCG
            721 GANGCCGTCC GGGGCCGGCG CGGCTCGCGC TCCCTCTCCG GCTCGGANTG CGAGGCAGCC
            781 CGCTCTCTTG CGCGCCTTGG CGGGCTCCTC TCCCTTCTGG AACCGANGCC CGGGCCGGGG
            841 CCTGNCTGCC GCTCGGCTGC CCCAAGCCTC TGCGCTNANC TTAAGTTTAA ACGCTAGNNA
            901 GCTTGGGNCT CCCTATAGTG ANNNNTANTN NNTTCNANNA NCNNGTNNNC ANNGGGNNCT
          961 CTANNNAGCN NNANANCTCT GCNNNNNNNN NN
          //
          Primer: hBIG2_F2 (Pr1020)
          Region: insert 3'
          Sequence file: RDB15365_A7I5e.seq check
          >D03588D2_A7I5_2_hBIG2_F2_F02_06_ABI08.ab1
              1 NNNNNNNNNN NGNNNNNNNN NNNNNTTCNN NAANNAGAAA CCNGCAGCCT GGCCTGTTGT
             61 TTGAGGATCC TGTTTCGAAT GTATGTTGAT GAGAACCGCA GGGATTCCTG GGAAGAAATA
            121 CAGCAGAGAC TTTTAACTGT TTGCAGTGAA GCTCTTGCCT ATTTCATCAC TGTGAATTCT
            181 GAGAGCCATC GGGAGGCCTG GACAAGTCTC TTGTTGTTAC TTCTAACTAA AACCCTTAAG
            241 ATAAATGATG AAAAGTTCAA AGCACATGCT TCAATGTACT ACCCCTACTT GTGTGAAATT
            301 ATGCAGTTTG ACCTGATCCC TGAGCTCCGA GCAGTTCTGC GGAAGTTCTT CCTACGGATA
            361 GGTGTTGTGT ATAAGATATG GATACCAGAA GAGCCATCAC AGGTACCAGC AGCACTGTCA
            421 CCAGTGTGGT AGCCCTGGCT GCCCAGGCCA GTGCTGCAGC TCTGCAGAAT GTTCAGCATG
            481 CCATTTCTGA CTGGCACATC TCGTGAAGTT TCATAGAAAC AAGGAGTTGG CATCTTGGAT
            541 CTCAGAATGG CCTGGAAACG GATGGCCTCT ACGCTGTTCC ATCACAGTCT CCAACTAAGG
            601 CTTATGGTAT TTCATTAAAC TGTTGCATAC CCAGTTAGCA CAGTAGGTGG GGAGTCTGCT
            661 TCATTTCTAT CATTCCATTT TTCTGATTAA ACTGTCAAAT CTGTCATTGC ATATGCCATC
            721 GTTTTCTAGC AAAATCCCAT GATTGNCTAT AAACGTTTTG TAAGAAGTCA CTCTCCTTGA
            781 AAATACTGAA CATAGCTGTA TAGGTTTGTG ATTATTAGAG AATATGTTAA TAAAACTTCT
            841 TGTAACGGCT AACTGCCACC TAAAATAAAA AAAAAAAAAA AAAACGTTNG CGGCNGCTCN
            901 ANNCTANNNG NNCCNNTTAA ACCCGCTGAN CANNCNNNNN NNNGNCNNTN CNANTTNGNN
          961 ANNNNTNNNN NNNNNN
          //
          Primer: SV40pro_F (Pr0110)
          Region: EM7 pro, ZeoR
          Sequence file: RDB15365_A7I5f.seq check
          >D03588D2_A7I5_2_SV40pro_F_D09_12_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNAN NNNTGNNGNN GCTTTTTTGG ANGCCTAGGC TTTTGCAAAA
             61 AGCTCCCGGG AGCTTGTATA TCCNTTTTCG GATCTGATCA GCACGTGTTG ACAATTAATC
            121 ATCGGCATAG TATATCGGCA TAGTATAATA CGACAAGGTG AGGAACTAAA CCATGGCCAA
            181 GTTGACCAGT GCCGTTCCGG TGCTCACCGC GCGCGACGTC GCCGGAGCGG TCGAGTTCTG
            241 GACCGACCGG CTCGGGTTCT CCCGGGACTT CGTGGAGGAC GACTTCGCCG GTGTGGTCCG
            301 GGACGACGTG ACCCTGTTCA TCAGCGCGGT CCAGGACCAG GTGGTGCCGG ACAACACCCT
            361 GGCCTGGGTG TGGGTGCGCG GCCTGGACGA GCTGTACGCC GAGTGGTCGG AGGTCGTGTC
            421 CACGAACTTC CGGGACGCCT CCGGGCCGGC CATGACCGAG ATCGGCGAGC AGCCGTGGGG
            481 GCGGGAGTTC GCCCTGCGCG ACCCGGCCGG CAACTGCGTG CACTTCGTGG CCGAGGAGCA
            541 GGACTGACAC GTGCTACGAG ATTTCGATTC CACCGCCGCC TTCTATGAAA GGTTGGGCTT
            601 CGGAATCGTT TTCCGGGACG CCGGCTGGAT GATCCTCCAG CGCGGGGNTC TCATGCTGGA
            661 GTTCTTCGCC CACCCCAACT TGTTTATTGC AGCTTATAAT GNTTACNAAT AAAGCAATAG
            721 CATCACAAAT TTCACAAATA AAGCATTTTT TTCACTGCAT TCTAGTTGTG GTTTGTCCNA
            781 ACTCATCAAT GTATCTTATC ATGTCTGTAT ACCGNCNNNC TCTANCTANA NNTNNNNGNN
            841 NNCATGGNCA TANNTGTTTC CNGNGNGAAN TNGNNNTCCG CNCACNNNTN CNNNNNNNNT
          901 ACNANNCNNN NNNNATAAAG NGNNANNNCN NGGGNNNNNN NANNNNNNNN NN
          //

          Please visit Sequencing and PCR primers for primer information.


          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.

          2023.05.01

          GNP_filter3_RDBDEP_html_230501.pl