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pET28/His6-EGFP-D4

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

Catalog number RDB13961
Resource name pET28/His6-EGFP-D4
Clone info. Marker of cholesterol-rich domain of the plasma membrane. Bacterial expression plasmid of His6-EGFP-tagged perfringolysin theta toxin D4 (domain 4 of theta toxin; K363-N472) of C. perfringens.
Comment Expression was confirmed by the depositor.
Vector backbone pET-28b (Plasmid)
Size of vector backbone 5.4 kb
Selectable markers Km^r
Gene/insert name Clostridium perfringens pfoA cDNA Aequorea victoria GFP cDNA
Depositor|Developer Kobayashi, Toshihide | Abe, Mitsuhiro |
Other clones in our bank
External Database
Clostridium perfringens pfoA Aequorea victoria GFP
          Reference sequence
            
           
          Sequence (full) RDB13961zzt01.seq compiled by us

          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(s) designated by the DEPOSITOR is requested. (Shimada, Y., Eur J Biochem. 269 (24): 6195-6203, 2002.)
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          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 (Shimada, Y., Eur J Biochem. 269 (24): 6195-6203, 2002.)。
          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。営利目的利用についてはお問い合わせください。
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          提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]

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          RDB13961 pET28/His6-EGFP-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-EGFP-D4 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13961).

          Reference section:

          Abe, M., Makino, A., Hullin-Matsuda, F., Kamijo, K., Ohno-Iwashita, Y., Hanada, K., Mizuno, H., Miyawaki, A., Kobayashi, T., A role for sphingomyelin-rich lipid domains in the accumulation of phosphatidylinositol-4,5-bisphosphate to the cleavage furrow during cytokinesis. Mol. Cell. Biol. 32 (8): 1396-1407 (2012). PMID 22331463.
          Shimada, Y., Maruya, M., Iwashita, S., Ohno-Iwashita, Y., The C-terminal domain of perfringolysin O is an essential cholesterol-binding unit targeting to cholesterol-rich microdomains. Eur. J. Biochem. 269 (24): 6195-6203 (2002). PMID 12473115.

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

          References and tips

          Featured content

          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 RDB13961_A6Btp1-4.pdf check

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

          Primer: pBR322_BamHI_R (Pr0065)
          Region: lacO,6xHis,thrombin site,insert 5'(EGFP 5')
          Sequence file: RDB13961_A6Btd.seq check
          >D03144A4_A6Bt_1_pBR 322 BamHI R_B04_04.ab1
              1 NNNNNNNNNN NNNNNTNNNN ATAGGNNNNN TGTGAGCGGA NAACAATTCC CCTCTAGAAA
             61 TAATTTTGTT TAACTTTAAG AAGGAGATAT ACCATGGGCA GCAGCCATCA TCATCATCAT
            121 CACAGCAGCG GCCTGGTGCC GCGCGGCAGC CATATGGCTA GAGTGAGCAA GGGCGAGGAG
            181 CTGTTCACCG GGGTGGTGCC CATCCTGGTC GAGCTGGACG GCGACGTAAA CGGCCACAAG
            241 TTCAGCGTGT CCGGCGAGGG CGAGGGCGAT GCCACCTACG GCAAGCTGAC CCTGAAGTTC
            301 ATCTGCACCA CCGGCAAGCT GCCCGTGCCC TGGCCCACCC TCGTGACCAC CCTGACCTAC
            361 GGCGTGCAGT GCTTCAGCCG CTACCCCGAC CACATGAAGC AGCACGACTT CTTCAAGTCC
            421 GCCATGCCCG AAGGCTACGT CCAGGAGCGC ACCATCTTCT TCAAGGACGA CGGCAACTAC
            481 AAGACCCGCG CCGAGGTGAA GTTCGAGGGC GACACCCTGG TGAACCGCAT CGAGCTGAAG
            541 GGCATCGACT TCAAGGAGGA CGGCAACATC CTGGGGCACA AGCTGGAGTA CAACTACAAC
            601 AGCCACAACG TCTATATCAT GGCCGACAAG CAGAAGAACG GCATCAAGGT GAACTTCAAG
            661 ATCCGCCACA ACATCGAGGA CGGCAGCGTG CAGCTCGCCG ACCACTACCA GCAGAACACC
            721 CCCATCGGCG ACNGCCCCGT GCTGCTGCCC GACAACCACT ACCTGANCAC CCAGTCCGCC
            781 CTGAGCAAAG ACCCCAACGA GAAGCGCGAT CACATGGTCC TGCTGGANTT CGTGACCGCC
            841 GCCNGGNTCA NTCTCGGCAT GGNCGANCTG TACNAGGCTA GCNAGGGAAA AATAAACTTA
            901 NNTCATAGTG NNNCCTATGT TGCNCANTTT GNNNNANCCN NGGNNTNANG TTTNNTATGA
          961 NNNNNNNNNA ANNNNANNTT TNNNNNNNNN AANNNNNGNN NNGNNANNNN NNNNNN
          //
          Primer: T7 terminator (Pr0155)
          Region: insert 3'(D4,EGFP 3')
          Sequence file: RDB13961_A6Bte.seq check
          >D03144A4_A6Bt_1_T7Terminator_A04_01.ab1
              1 NNNNNNNNNN TNNNNNCTTN NTNNGCNGCC GGNNCTCNGT GGTGGTGGTG GTGGTGCTCG
             61 AGTGCGGCCG CAANCTTGTC GACGGAGCTC GAATTCGAGC TCGGTACCCT ATGAATCTAA
            121 TACATGTAAA CCAAATTTAT TGTATAGTAA TGTCCAAAAC AAATCTAATA TTTACTTTAT
            181 TATAACTCTA AAATATGAAA TAAAAAACTG ATTTTAGACT AGCTAAAATC AGTTTTTACT
            241 TTAGTTTAAT TGTAAGTAAT ACTAGATCCA GGGTATAAAG TTGTTCCCCA TATTGAAACA
            301 TTTATATTAT TTGTTAATGG AACATCATAT TCACTTATAA CATCTCTCCA CCATTCCCAA
            361 GCAAGGCCTG TACACTCTCT TGCTTTTATT CTTATATTTC TTGCATTAGC TTCAAGAGGT
            421 ATTACTGTTG AATAGTGAGC TGTTTTATCT TGATAATTTC CATCCCATGT TTTATGAGTT
            481 AAAACTTCAT TTCCTTCTTT GTCATATGAA ACTTCATCCC AGGCTACTTC AAACTGTGCA
            541 ACATAGGCTC CACTATGATC TAAGTTTATT TTTCCCTTGC TAGCCTTGTA CAGCTCGTCC
            601 ATGCCGAGAG TGATCCCGGC GGCGGTCACG AACTCCAGCA GGACCATGTG ATCGCGCTTC
            661 TCGTTGGGGT CTTTGCTCAN GGCGGANTGG GTGCTCAGGT AGTGGTTGTC GGGCAGCAGC
            721 ACGGGGCCGT CGCCGATGGG GGTGTTCTGC TGGTAGTGGT CGGNGAGCTG CACGCTGCCG
            781 TCCTCGATGT TGTGGCGGNT CTTGAAGTTC ACCTTGATGC CGTTCTTCTG CTTGTCNGCC
            841 ATGATATANA CGTTGTGNNT GTTGTAGTTG TACTCCNGCT NNNGCCCCNN NNGNTGCCNN
            901 CCNNNNNNNA GNCNATGCCC TTCAGCTCNA TGNNNNNCNN NANGGNGNCG CCNNCNAACT
          961 TCNNNNNNNC NCGGGNNNNN NNNNN
          //
          Primer: EGFP-N_F2 (Pr1228)
          Region: insert mid(EGFP 3',D4 5')
          Sequence file: RDB13961_A6Btf.seq check
          >13961_B3DpF_EGFP-N_F2_B03_02_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNC NNNNNNNNNG ANGGCAACTA CNNNACCCGN
             61 NNNNAGGTGA AGTTCNAGNN NNNCACCCTG GTGAACCGCA TCGAGCTGAA GGGCATCNNN
            121 TTCAAGGAGG ACGGCAACAT CCTGGGGCAC AAGCTGGAGT ACAACTACNN CAGCCACAAC
            181 GTCTATATCA TGGCCGACAA GCAGAAGAAC GGCATCAAGG TGAACTTCAA GATCCGCCAC
            241 AACATCGAGG ACGGCAGCGT GCAGCTCGCC GACCACTACC AGCAGAACAC CCCCATCGGC
            301 GACGGCCCCG TGCTGCTGCC CGACAACCAC TACCTGAGCA CCCAGTCCGC CCTGAGCAAA
            361 GACCCCAACG AGAAGCGCGA TCACATGGTC CTGCTGGAGT TCGTGACCGC CGCCGGGATC
            421 ACTCTCGGCA TGGACGAGCT GTACAAGGCT AGCAAGGGAA AAATNNNNNN NGATCATAGT
            481 GGAGCCTATG TTGCACAGTT TGAAGTAGCC TGGGATGAAG TTTCATATGA CAAAGAAGGA
            541 AATGAAGTTT TAACTCATAA AACATGGGAT GGAAATTATC AAGATAAAAC AGCTCACTAT
            601 TCAACAGTAA TACCTCTTGA AGCTAATGCA AGAAATATAA GAATAAAAGC AAGAGAGTGT
            661 ACAGGCCTTG CTTGGGAATG GTGNANAGAT GTTATNNNNN AATNNNNNNT TCCATTAACN
            721 AAATAATATA AATGTTTCAA TATGGGGAAC AACTTTATAC CCTGGATCTA GTATTACTTA
            781 CAATTNAANC TAAAGTAAAA ACTGATTTTA GCTNNNTCTA AAATCAGTTT TTTANTTCAT
            841 ATTTTANAAN NTTATAATAA AGTAAATATT AGANTTTGNT TTTGGACANN NACTATACNA
            901 NTAAANTTTN GGNTTACNTG TATNANANTT CNTANGGGTN NCCNANCNNN AATTCNAGCT
            961 CCNTCNNNAA GCTNGCNGGC CGNNCTCNAN CANCNNNCCN NNNNCACCNC TGANANCNGG
           1021 CTGNNNNANN AANNNCNNAA ANGNANGNTG NANTTGGNNN GNNNGNCNNN CNNNNNAANC
          1081 NAATNNNNNN NNNN
          //
          Primer: pGL3_RV4 (Pr0335)
          Region: lacI 3'
          Sequence file: RDB13961_A6Btg.seq check
          >D03144A4_A6Bt_1_pGL3_RV4_F12_18.ab1
              1 NNNNNNNNNN NNNNNNGNTC TCAGGGCNTC GGTCGAGNTC CCGGTGCCTA ATGAGTGAGC
             61 TAACTTACAT TAATTGCGTT GCGCTCACTG CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC
            121 CAGCTGCATT AATGAATCGG CCAACGCGCG GGGAGAGGCG GTTTGCGTAT TGGGCGCCAG
            181 GGTGGTTTTT CTTTTCACCA GTGAGACGGG CAACAGCTGA TTGCCCTTCA CCGCCTGGCC
            241 CTGAGAGAGT TGCAGCAAGC GGTCCACGCT GGTTTGCCCC AGCAGGCGAA AATCCTGTTT
            301 GATGGTGGTT AACGGCGGGA TATAACATGA GCTGTCTTCG GTATCGTCGT ATCCCACTAC
            361 CGAGATATCC GCACCAACGC GCAGCCCGGA CTCGGTAATG GCGCGCATTG CGCCCAGCGC
            421 CATCTGATCG TTGGCAACCA GCATCGCAGT GGGAACGATG CCCTCATTCA GCATTTGCAT
            481 GGTTTGTTGA AAACCGGACA TGGCACTCCA GTCGCCTTCC CGTTCCGCTA TCGGCTGAAT
            541 TTGATTGCGA GTGAGATATT TATGCCAGCC AGCCAGACGC AGACGCGCCG AGACAGAACT
            601 TAATGGGCCC GCTAACAGCG CGATTTGCTG GTGACCCAAT GCGACCAGAT GCTCCACGCC
            661 CAGTCGCGTA CCGTCTTCAT GGGAGAAAAT AATACTGTTG ATGGGTGTCT GGNCAGAGAC
            721 ATCAAGAAAT AACGCCNGAA CATTAGTGCA NGCAGCTTCC ACAGCAATGG CATCCTGGTC
            781 ATCCAGCGGA TAGTTAATGA NCAGCCCACT GACGCGTTGC GCGAGAAGAN TGTGCNNCGC
            841 CGCTTTACAN GNTTCNACGC CGCTTCNTTC NACCATCNNN ACCACCNCGC NNNNNNCCAG
          901 TTNNTCNGCG CGANANTTNA TCNNNNCGAN NNTTTNCNNN NNNNNNN
          //

          Please visit Sequencing and PCR primers for primer information.

          References

          Original, user report and related articles

          original Abe, M., A role for sphingomyelin-rich lipid domains in the accumulation of phosphatidylinositol-4,5-bisphosphate to the cleavage furrow during cytokinesis. Mol. Cell. Biol. 32 (8): 1396-1407 (2012). PMID 22331463.
          original Shimada, Y., The C-terminal domain of perfringolysin O is an essential cholesterol-binding unit targeting to cholesterol-rich microdomains. Eur. J. Biochem. 269 (24): 6195-6203 (2002). PMID 12473115.
          reference Abe, M., Imaging cholesterol depletion at the plasma membrane by methyl-β-cyclodextrin. J. Lipid Res. 62:100077 (2021). PMID 33891936.
          protocol Abe, M., Dynamics of sphingomyelin-and cholesterol-enriched lipid domains during cytokinesis. Methods Cell Biol. 137: 15-24 (2017). PMID 28065303.
          user_report Lee, J.H., 25-Hydroxycholesterol suppress IFN-γ-induced inflammation in microglia by disrupting lipid raft formation and caveolin-mediated signaling endosomes. Free Radic. Biol. Med. 179: 252-265 (2022). PMID 34808332.

          2023.05.22

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