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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-EGFP-NT-Lys

Marker of sphingomyelin in cell membrane. Bacterial expression plasmid of nontoxic Lysenin.

Catalog number RDB13498
Resource name pET28/His6-EGFP-NT-Lys
Clone info. Marker of sphingomyelin in cell membrane. Bacterial expression plasmid of nontoxic Lysenin.
Comment 169-297 AA mutant. Reference of DNA sequence, D85846.1.
Vector backbone pET-28b (Plasmid)
Size of vector backbone 5.4 kb
Selectable markers Kan^r
Gene/insert name common brandling worm Lysenin cDNA Aequorea victoria GFP cDNA
Depositor|Developer Kobayashi, Toshihide | Abe, Mitsuhiro |
Other clones in our bank
External Database
common brandling worm Lysenin
    Aequorea victoria GFP
            Reference sequence
              
             
            Sequence (full) RDB13498zzt01.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 designated by the DEPOSITOR is requested. (Makino, A., FASEB J. 29 (2): 477-493, 2015)
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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(Makino, A., FASEB J. 29 (2): 477-493, 2015)。
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            手続きの概要は、「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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            RDB13498 pET28/His6-EGFP-NT-Lys 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-NT-Lys was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13498).

            Reference section:

            Makino, A., Abe, M., Murate, M., Inaba, T., Yilmaz, N., Hullin-Matsuda, F., Kishimoto, T., Schieber, N.L., Taguchi, T., Arai, H., Anderluh, G., Parton, R.G., Kobayashi, T., Visualization of the heterogeneous membrane distribution of sphingomyelin associated with cytokinesis, cell polarity, and sphingolipidosis. FASEB J. 29 (2): 477-493 (2015). PMID 25389132. [link to RRC of NBRP]

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

            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 RDB13498_A6Cap1-2.pdf check

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

            Primer: T7_20bp (Pr0261)
            Region: 6xHis,thrombin site,insert 5'(EGFP 5')
            Sequence file: RDB13498_A6Cad.seq check
            >13498_13498_A6Ca_4_T7_20bp_E05_14.ab1
                1 NNNNNNNNNN NNNNNNNNNN NTTTGTTNNN CTTTAAGAAG GAGNNATACC ATGGGCAGCA
               61 GCCATCATCA TCATCATCAC AGCAGCGGCC TGGTGCCGCG CGGCAGCCAT ATGGCTAGAG
              121 TGAGCAAGGG CGAGGAGCTG TTCACCGGGG TGGTGCCCAT CCTGGTCGAG CTGGACGGCG
              181 ACGTAAACGG CCACAAGTTC AGCGTGTCCG GCGAGGGCGA GGGCGATGCC ACCTACGGCA
              241 AGCTGACCCT GAAGTTCATC TGCACCACCG GCAAGCTGCC CGTGCCCTGG CCCACCCTCG
              301 TGACCACCCT GACCTACGGC GTGCAGTGCT TCAGCCGCTA CCCCGACCAC ATGAAGCAGC
              361 ACGACTTCTT CAAGTCCGCC ATGCCCGAAG GCTACGTCCA GGAGCGCACC ATCTTCTTCA
              421 AGGACGACGG CAACTACAAG ACCCGCGCCG AGGTGAAGTT CGAGGGCGAC ACCCTGGTGA
              481 ACCGCATCGA GCTGAAGGGC ATCGACTTCA AGGAGGACGG CAACATCCTG GGGCACAAGC
              541 TGGAGTACAA CTACAACAGC CACAACGTCT ATATCATGGC CGACAAGCAG AAGAACGGCA
              601 TCAAGGTGAA CTTCAAGATC CGCCACAACA TCGAGGACGG CAGCGTGCAG CTCGCCGACC
              661 ACTACCAGCA GAACACCCCC ATCGGCGACG GCCCCGTGCT GCTGCCCGAC AACCACTACC
              721 TGANCACCCA GTCCGCCCTG ANCAAAGANC CCAACGAGAA GCGCGATCAC ATGGTCCTGC
              781 TGGANTTCGT GACCGCCGCC GGGATCACTC TCGGCATGNA CGANCTGTAC AAGAAGCTTA
              841 TAATCCTTGG AAAANCAGAA NNTCGAANTN AGCANGCAGA AANGAANGNN TACATGACAG
              901 TCGTTNCNAG AAAAAAGTTN GCCAGCNGCN NNNNTTGGNN ATANCAAACT TTTTCNNNNT
            961 NGNGNNNNAN NNNNNNNNN
            //
            Primer: T7 terminator (Pr0155)
            Region: insert 3'(NT-Lys, EGFP 3')
            Sequence file: RDB13498_A6Cae.seq check
            >13498_13498_A6Ca_4_T7_terminator_F05_17.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNTNAGCAGC CGGNNCTCAG TGGTGGTGGT GGTGGTGCTC
               61 GAGTTAACCA ACCACTTCCA AAATCCACTT GTCTTCACGT TTGTCCAGAC AGTACACGTT
              121 TGTTGCCGGT CCATCATCGT AGCACAGCCC ACTGCGAGTG AAGTACTTAT TCATGAAGGT
              181 GACTACGTCA CCATGACGAA GAGGCAATGA CTTATTGATT GCCCAGCGCT GTTTCGGATT
              241 ATCAGTACCC TGATCAAAGT AGATTCCTCC TTGATCAGAG GAATAGGCAT ACTCATAGCC
              301 CGAATACATG GTGTTCAGCG TTTTAATTCG AAATCCCCCC CAATCTTCAT AGAGCACAAA
              361 CTTGAAAAGT TTGCTATGTC CAAGAGTTGC AGCTGGCCAA CTTTTTCTTG AAACGACTGT
              421 CATGTACTCC TTCCTTTCTG CATGCTTAAT TCGAATTTCT GTTTTTCCAA GGATTATAAG
              481 CTTCTTGTAC AGCTCGTCCA TGCCGAGAGT GATCCCGGCG GCGGTCACGA ACTCCAGCAG
              541 GACCATGTGA TCGCGCTTCT CGTTGGGGTC TTTGCTCAGG GCGGACTGGG TGCTCAGGTA
              601 GTGGTTGTCG GGCAGCAGCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT GGTAGTGGTC
              661 GGCGAGCTGC ACGCTGCCGT CCTCGATGTT GTGGCGGATC TTGAAGTTCA CCTTGATGCC
              721 GTTCTTCTGC TTGTCGGCCA TGATATAGAC GTTGTGGCTG TTGTAGTTGT ACTCCAGCTT
              781 GTGCCCCANG ATGTTGCCGT CCTCCTTGAA GTCNATGCNN TTCAGCTCGA TGCGGTTCAC
              841 CAGGGTGTCG CCCTCGAACT TCACNTCGGC GCNGGTCNTG TAGTTGNCGT CGTNCNNNAA
              901 GAANANGGTG CGCNNCNNGN NGTANCCNNC NGGNNTGGNG GNNTNGNANN AANNCGTGCT
            961 GCTNNNTGTN NNCGGGNNAN CGGNNGNNNN NNNNNN
            //
            Primer: T7 terminator (Pr0155)
            Region: insert 3'(NT-Lys, EGFP 3')
            Sequence file: RDB13498_A6Cae.seq check
            >13498_13498_A6Ca_4_T7_terminator_F05_17.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNTNAGCAGC CGGNNCTCAG TGGTGGTGGT GGTGGTGCTC
               61 GAGTTAACCA ACCACTTCCA AAATCCACTT GTCTTCACGT TTGTCCAGAC AGTACACGTT
              121 TGTTGCCGGT CCATCATCGT AGCACAGCCC ACTGCGAGTG AAGTACTTAT TCATGAAGGT
              181 GACTACGTCA CCATGACGAA GAGGCAATGA CTTATTGATT GCCCAGCGCT GTTTCGGATT
              241 ATCAGTACCC TGATCAAAGT AGATTCCTCC TTGATCAGAG GAATAGGCAT ACTCATAGCC
              301 CGAATACATG GTGTTCAGCG TTTTAATTCG AAATCCCCCC CAATCTTCAT AGAGCACAAA
              361 CTTGAAAAGT TTGCTATGTC CAAGAGTTGC AGCTGGCCAA CTTTTTCTTG AAACGACTGT
              421 CATGTACTCC TTCCTTTCTG CATGCTTAAT TCGAATTTCT GTTTTTCCAA GGATTATAAG
              481 CTTCTTGTAC AGCTCGTCCA TGCCGAGAGT GATCCCGGCG GCGGTCACGA ACTCCAGCAG
              541 GACCATGTGA TCGCGCTTCT CGTTGGGGTC TTTGCTCAGG GCGGACTGGG TGCTCAGGTA
              601 GTGGTTGTCG GGCAGCAGCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT GGTAGTGGTC
              661 GGCGAGCTGC ACGCTGCCGT CCTCGATGTT GTGGCGGATC TTGAAGTTCA CCTTGATGCC
              721 GTTCTTCTGC TTGTCGGCCA TGATATAGAC GTTGTGGCTG TTGTAGTTGT ACTCCAGCTT
              781 GTGCCCCANG ATGTTGCCGT CCTCCTTGAA GTCNATGCNN TTCAGCTCGA TGCGGTTCAC
              841 CAGGGTGTCG CCCTCGAACT TCACNTCGGC GCNGGTCNTG TAGTTGNCGT CGTNCNNNAA
              901 GAANANGGTG CGCNNCNNGN NGTANCCNNC NGGNNTGGNG GNNTNGNANN AANNCGTGCT
            961 GCTNNNTGTN NNCGGGNNAN CGGNNGNNNN NNNNNN
            //

            Please visit Sequencing and PCR primers for primer information.

            References

            Original, user report and related articles

            original Makino, A., Visualization of the heterogeneous membrane distribution of sphingomyelin associated with cytokinesis, cell polarity, and sphingolipidosis. FASEB J. 29 (2): 477-493 (2015). PMID 25389132. [link to RRC of NBRP]
            protocol Abe, M., Dynamics of sphingomyelin-and cholesterol-enriched lipid domains during cytokinesis. Methods Cell Biol. 137: 15-24 (2017). PMID 28065303.
            review Kobayashi, T., Impact of Intrinsic and Extrinsic Factors on Cellular Sphingomyelin Imaging with Specific Reporter Proteins.
            user_report Liang, H., The formation of migrasomes is initiated by the assembly of sphingomyelin synthase 2 foci at the leading edge of migrating cells. Nat. Cell Biol. 25 (8): 1173-1184 (2023). PMID 37488437. [link to RRC of NBRP]
            user_report Mori, Y., Membrane Sphingomyelin in Host Cells Is Essential for Nucleocapsid Penetration into the Cytoplasm after Hemifusion during Rubella Virus Entry. mBio 13 (6): e0169822 (2022). PMID 36346228. [link to RRC of NBRP]

            2023.09.13

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