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

pET28/His6-mCherry-D4

Marker of cholesterol-rich domain of the plasma membrane. Bacterial expression plasmid of His6-mCherry-tagged perfringolysin theta toxin D4 of C. perfringens.

Catalog number RDB14300
Resource name pET28/His6-mCherry-D4
Clone info. Marker of cholesterol-rich domain of the plasma membrane. Bacterial expression plasmid of His6-mCherry-tagged perfringolysin theta toxin D4 (domain 4 of theta toxin; K363-N472) of C. perfringens, see Fig. 1A of Shimada, Y. et. al. Eur. J. Biochem, 269: 6195-6203, 2002.
Comment Expression was confirmed by the depositor with SDS-PAGE. DQ673099.1 for Clostridium perfringens CN3947 perfringolysin O (pfoA) gene.
Vector backbone pET-28b (Plasmid)
Size of vector backbone 5.4 kb
Selectable markers Kan^r
Gene/insert name Clostridium perfringens perfringolysin O (pfoA) cDNA
Depositor|Developer Sako, Yasushi | Abe, Mitsuhiro |
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External Database
Clostridium perfringens perfringolysin O (pfoA)

          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 (Makino, A. et al., FASEB J. 31 (4): 1301-1322 (2017)).
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          Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
          Please visit Information of Request for Distribution.[link] 
          Remarks
          This RESOURCE contains a fluorescent protein provided by Prof. Roger Tsien. UCSD's MTA is also required. Please visit Additional Forms section. This RESOURCE is not provided to for-profit organization or for for-profit research by non-profit organizations.
          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 (Makino, A. et al., FASEB J. 31 (4): 1301-1322 (2017))。
          提供依頼
          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]
          手続きの概要は、「提供申込みについて[link]」をご覧ください。
          備考
          Prof. Roger Tsienから供与された蛍光タンパク質を含みます。UCSDとのMTA締結が必要です。「申込みに必要な書式」をご覧ください。本件リソースは営利機関および非営利機関による営利目的研究には提供いたしません。

          Catalog # Resource name Shipping form Fee (non-profit org.)
          RDB14300 pET28/His6-mCherry-D4 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 pET28/His6-mCherry-D4 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB14300).

          Reference section:

          Makino, A., Abe, M., Ishitsuka, R., Murate, M., Kishimoto, T., Sakai, S., Hullin-Matsuda, F., Shimada, Y., Inaba, T., Miyatake, H., Tanaka, H., Kurahashi, A., Pack, C.G., Kasai, R.S., Kubo, S., Schieber, N.L., Dohmae, N., Tochio, N., Hagiwara, K., Sasaki, Y., Aida, Y., Fujimori, F., Kigawa, T., Nishibori, K., Parton R.G., Kusumi, A., Sako, Y., Anderluh, G., Yamashita, M., Kobayashi, T., Greimel, P., Kobayashi, T., A novel sphingomyelin/cholesterol domain-specific probe reveals the dynamics of the membrane domains during virus release and in Niemann-Pick type C. FASEB J. 31 (4): 1301-1322 (2017). PMID 27492925. [link to RRC of NBRP]

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


          References and tips

          Featured content

          Featured content Dnaconda's recommendation EXR0062e (English text)
          Featured content Dnaconda's recommendation EXR0062j (Japanese text)
          Featured content Visualization of Organelles (English text)
          Featured content Sphingolipid signaling pathway (English text)

          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 RDB14300_A6Hup1-4.pdf check

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

          Primer: T7_20bp (Pr0261)
          Region: 6xHis,thrombin site,insert 5' (mCherry 5')
          Sequence file: RDB14300_A6Hud.seq check
          >D03762A1_A6Hu_3_T7_20bp_A02_01_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NTTNNNNNNN NNNNNNNNNA GGAGANATAC
             61 CATGGGCAGC AGCCATCATC ATCATCATCA CAGCAGCGGC CTGGTGCCGC GCGGCAGCGG
            121 TACCATGGTG AGCAAGGGCG AGGAGGATAA CATGGCCATC ATCAAGGAGT TCATGCGCTT
            181 CAAGGTGCAC ATGGAGGGCT CCGTGAACGG CCACGAGTTC GAGATCGAGG GCGAGGGCGA
            241 GGGCCGCCCC TACGAGGGCA CCCAGACCGC CAAGCTGAAG GTGACCAAGG GTGGCCCCCT
            301 GCCCTTCGCC TGGGACATCC TGTCCCCTCA GTTCATGTAC GGCTCCAAGG CCTACGTGAA
            361 GCACCCCGCC GACATCCCCG ACTACTTGAA GCTGTCCTTC CCCGAGGGCT TCAAGTGGGA
            421 GCGCGTGATG AACTTCGAGG ACGGCGGCGT GGTGACCGTG ACCCAGGACT CCTCCCTGCA
            481 GGACGGCGAG TTCATCTACA AGGTGAAGCT GCGCGGCACC AACTTCCCCT CCGACGGCCC
            541 CGTAATGCAG AAGAAGACCA TGGGCTGGGA GGCCTCCTCC GAGCGGATGT ACCCCGAGGA
            601 CGGCGCCCTG AAGGGCGAGA TCAAGCAGAG GCTGAAGCTG AAGGACGGCG GCCACTACGA
            661 CGCTGAGGTC AAGACCACCT ACNAGGCCAA GAAGCCCGTG CAGCTGCCCG GCGCCTACAA
            721 CGTCAACATC AAGTTGGACA TCACCTCCCA CNANCGAGGA CTACACCATC GTGGAACAGT
            781 ACGAACGCGC CGAGGGCCGC CNCTCCANCG GCGGCATGGA CNAGCTGTAC AAGANNCTTA
            841 AGGGAAAAAT NAACTTAGAT CATANTGGAG CCTATGNTGC ACAGTTTGAA GTANCNTGGN
            901 NNNGANGTTT CATATGACNA AGANNNAATG AANNTTTANN TCNTAAACAT GGGATGNNAA
            961 TTATCNNGAN AANACAGCTC ACTANTNNNA CNNTNNNNCC NCNTGAANCT NATGNNANNN
          1021 NNNNAANNNN NAANNCNNGA NNNNNNNCNN NNCNNNNCNT NNNNNNNNGN NNNNNNNNN
          //
          Primer: T7 terminator (Pr0155)
          Region: insert 3'(D4,mCherry 3')
          Sequence file: RDB14300_A6Hue.seq check
          >D03762A1_A6Hu_3_T7 Terminator_B02_02_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNGNTTNNTN NGCAGCCGGN NCTCNGTGGT GGTGGTGGTG
             61 GTGCTCGAGT TAATTGTAAG TAATACTAGA TCCAGGGTAT AAAGTTGTTC CCCATATTGA
            121 AACATTTATA TTATTTGTTA ATGGAACATC ATATTCACTT ATAACATCTC TCCACCATTC
            181 CCAAGCAAGG CCTGTACACT CTCTTGCTTT TATTCTTATA TTTCTTGCAT TAGCTTCAAG
            241 AGGTATTACT GTTGAATAGT GAGCTGTTTT ATCTTGATAA TTTCCATCCC ATGTTTTATG
            301 AGTTAAAACT TCATTTCCTT CTTTGTCATA TGAAACTTCA TCCCAGGCTA CTTCAAACTG
            361 TGCAACATAG GCTCCACTAT GATCTAAGTT TATTTTTCCC TTAAGCTTCT TGTACAGCTC
            421 GTCCATGCCG CCGGTGGAGT GGCGGCCCTC GGCGCGTTCG TACTGTTCCA CGATGGTGTA
            481 GTCCTCGTTG TGGGAGGTGA TGTCCAACTT GATGTTGACG TTGTAGGCGC CGGGCAGCTG
            541 CACGGGCTTC TTGGCCTTGT AGGTGGTCTT GACCTCAGCG TCGTAGTGGC CGCCGTCCTT
            601 CAGCTTCAGC CTCTGCTTGA TCTCGCCCTT CAGGGCGCCG TCCTCGGGGT ACATCCGCTC
            661 GGAGGAGGCC TCCCAGCCCA TGGTCTTCTT CTGCATTACG GGGCCGTCGG ANGGGAAGTT
            721 GGTGCCGCGC AGCTTCACCT TGTAGATGAA CTCGCCGTCC TGCANGGANG ANTCCTGGGT
            781 CACGGTCACC ACGCCGCCGT CCTCGAAGTT CATCACGCGC TCCCACTTGA AGNNNTCGGG
            841 GAAGGACAGC TTCAAGTANT CGGGGANGTC NGCGGGGNGN TTCACGTANG CNTTNGNANN
            901 NGTACATGAN CTNANGGGGA NAGGANGTCC CANNCGAANG NNANGGGGCC ACCCTNNGNC
          961 ANNNNCNNCN TNNNNNNCNG GGNGNCCNNN NNNGNNNNNN NNNN
          //
          Primer: mCherry-N_rev (Pr0398)
          Region: thrombin site,6x His,RBS,lacO, T7 pro,lacI pro,lacI
          Sequence file: RDB14300_A6Huf.seq check
          >D03762A1_A6Hu_3_mCherry-N_rev_A02_02_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNGN NGGNNNTGTT ANCCTCCTCG CCCTTGCTCN CCATGGTACC
             61 GCTGCCGCGC GGNACCAGGC CGCTGCTGTG ATGATGATGA TGATGGCTGC TGCCCATGGT
            121 ATATCTCCTT CTTAAAGTTA AACAAAATTA TTTCTAGAGG GGAATTGTTA TCCGCTCACA
            181 ATTCCCCTAT AGTGAGTCGT ATTAATTTCG CGGGATCGAG ATCTCGATCC TCTACGCCGG
            241 ACGCATCGTG GCCGGCATCA CCGGCGCCAC AGGTGCGGTT GCTGGCGCCT ATATCGCCGA
            301 CATCACCGAT GGGGAAGATC GGGCTCGCCA CTTCGGGCTC ATGAGCGCTT GTTTCGGCGT
            361 GGGTATGGTG GCAGGCCCCG TGGCCGGGGG ACTGTTGGGC GCCATCTCCT TGCATGCACC
            421 ATTCCTTGCG GCGGCGGTGC TCAACGGCCT CAACCTACTA CTGGGCTGCT TCCTAATGCA
            481 GGAGTCGCAT AAGGGAGAGC GTCGAGATCC CGGACACCAT CGAATGGCGC AAAACCTTTC
            541 GCGGTATGGC ATGATAGCGC CCGGAAGAGA GTCAATTCAG GGTGGTGAAT GTGAAACCAG
            601 TAACGTTATA CGATGTCGCA GAGTATGCCG GTGTCTCTTA TCAGACCGTT TCCCGCGTGG
            661 TGAACCAGGC CAGCCACGTT TCTGCGAAAA CGCGGGAAAA AGTGGAAGCG GCGATGGCGG
            721 AGCTGANTTA CATTCCCAAC CGCGTGGCAC AACAACTGGC GGGCAAACAG TCGTTGCTGA
            781 TTGGCGTTGC CACATCCAGT CTGGCCCTGC ACGCGCCGTC GCAAATTGTC GCGGCGATTA
            841 AATCTCGCGC CGATCAACTG GGTGCCAGCG NGGTGGNGTC GATGGNAGAA CGAAGCGGCG
            901 TCNAAGCCTG NAAAGCGGCG GNGCACAATC TTCTCGCGCA ACGCGNCAGN NGGCTGATCN
          961 TNNANNATCC NCNNGNATNA NNNNNNNGNN NNNNNNNN
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

          original Makino, A., A novel sphingomyelin/cholesterol domain-specific probe reveals the dynamics of the membrane domains during virus release and in Niemann-Pick type C. FASEB J. 31 (4): 1301-1322 (2017). PMID 27492925. [link to RRC of NBRP]
          reference Abe, M., Imaging cholesterol depletion at the plasma membrane by methyl-β-cyclodextrin. J. Lipid Res. 62:100077 (2021). PMID 33891936.
          user_report Miyata, Y., Protocol to analyze lipid asymmetry in the plasma membrane. STAR Protoc. 3 (4): 101870 (2022). PMID 36595929. [link to RRC of NBRP]
          user_report Kitano, V.J.F., LDL uptake-dependent phosphatidylethanolamine translocation to the cell surface promotes fusion of osteoclast-like cells. J. Cell Sci. 133 (10): jcs243840 (2020). PMID 32295848. [link to RRC of NBRP]

          2023.06.20

          GNP_filter3_RDBDEP_html_230615.pl