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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.
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 Kobayashi, Toshihide | Abe, Mitsuhiro |

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 set forth by the DEPOSITOR 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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Schedule A (GFP-A) [Link]
Remarks
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RDB13961 pET28/His6-EGFP-D4 DNA solution

Ordering Information [in Japanese] [in English]

References and tips

Electronic file

Original reference

original Abe, M., Dynamics of sphingomyelin-and cholesterol-enriched lipid domains during cytokinesis. Methods Cell Biol. 137: 15-24 (2017). PMID 28065303.
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.

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

Featured content

Featured content Sphingolipid signaling pathway (English text)
Featured content Organelle marker (English text)

Sequence information

RIKEN BRC has sequenced portions of this material for quality test. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB13961_A6Btp1-2.pdf

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

Primer: pBR 322 BamHI R RDB13961_A6Bta.seq
>D03144A4_A6Bt_1_pBR 322 BamHI R_B04_04.ab1
    1 TCCGACATAT ACGACTCACT ATAGGGGAAT TGTGAGCGGA TAACAATTCC 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 ACGGCCCCGT GCTGCTGCCC GACAACCACT ACCTGAGCAC CCAGTCCGCC
  781 CTGAGCAAAG ACCCCAACGA GAAGCGCGAT CACATGGTCC TGCTGGAGTT CGTGACCGCC
  841 GCCGGGATCA CTCTCGGCAT GGACGAGCTG TACAAGGCTA GCAAGGGAAA AATAAACTTA
  901 GATCATAGTG GAGCCTATGT TGCACAGTTT GAAGTAGCCT GGGGATGAAG TTTCATATGA
  961 CAAGAATGGA AATGAAAGTT TTAACTCATA AAACATGGGA TGGGAAATTA TCAAGATAAA
 1021 ACAGCTCACT AATTCACAGT AATACCCTCT GAGGCTTAAT TGCAGAAATA ATAGAATAAG
1081 CCAGAGGACT GTACAGGTCC TTGGCTTTGG GGGCAAATGG TTTT
//
Primer: T7Terminator RDB13961_A6Btb.seq
>D03144A4_A6Bt_1_T7Terminator_A04_01.ab1
    1 CAATGCATCT TTCGGGCTTT GTTAGCAGCC GGATCTCAGT GGTGGTGGTG GTGGTGCTCG
   61 AGTGCGGCCG CAAGCTTGTC 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 CTTTGCTCAG GGCGGACTGG GTGCTCAGGT AGTGGTTGTC GGGCAGCAGC
721 ACGGGGCCGT CGCCGATGGG GGTGTTCTGC TGGTAGTGGT CGGCGAGCTG CACGC
//
Primer: pGL3_RV4 RDB13961_A6Btc.seq
>D03144A4_A6Bt_1_pGL3_RV4_F12_18.ab1
    1 GGGACGATGG GTTGAGGCTC TCAGGGCATC GGTCGAGATC 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 GGTCAGAGAC
  721 ATCAAGAAAT AACGCCGGAA CATTAGTGCA GGCAGCTTCC ACAGCAATGG CATCCTGGTC
  781 ATCCAGCGGA TAGTTAATGA TCAGCCCACT GACGCGTTGC GCGAGAAGAT TGTGCACCGC
  841 CGCTTTACAG GCTTCGACGC CGCTTCGTTC TACCATCGAC ACCACCACGC TGGCACCCAG
  901 TTGATCGGCG CGAGATTTAA TCGCCGCGAC AATTTGCGAC GGCGCGTGCA GGGCCAGACT
  961 GGAGTGGCAC GCCCATCAGC AACGACTGTT TGCCGCCAGT GTTGTGCCAC GCGTTGGGAT
 1021 GTAATCCAGC TCGCAATCGC CGGCTCAACT TTTTCCCGCG TTTCGCAGAA CGTTGTCTGA
1081 CTGGTCACCC ACGCGGGGAA CT
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Abe, M., Dynamics of sphingomyelin-and cholesterol-enriched lipid domains during cytokinesis. Methods Cell Biol. 137: 15-24 (2017). PMID 28065303.
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.

2019.06.21

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