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pET28/PlyA2-EGFP-His6 (#RDB13962)

Marker of sphingomyelin and cholesterol lipid domains. Bacterial expression plasmid of EGFP-His6-tagged PlyA2.

Clone info. Marker of sphingomyelin and cholesterol lipid domains. Bacterial expression plasmid of EGFP-His6-tagged PlyA2.
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 Pleurotus eryngii PlyA2 cDNA Aequorea victoria GFP cDNA
Depositor|Developer Kobayashi, Toshihide | Abe, Mitsuhiro |
 
Sequence (full) RDB13962zzt01.seq compiled by us

Distribution information

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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. (Bhat, HB., J Lipid Res. 54 (10): 2933-2943, 2013.)
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提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
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。
MTAに書く使用条件 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 (Bhat, HB., J Lipid Res. 54 (10): 2933-2943, 2013.)。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB13962 pET28/PlyA2-EGFP-His6 DNA solution JPY 9,460 (not-for-profit academic purpose)
plus cost of shipping containers, dry ice (if required) and shipping charge

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/PlyA2-EGFP-His6 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13962).

Reference section:

Bhat, H.B., Ishitsuka, R., Inaba, T., Murate, M., Abe, M., Makino, A., Kohyama-Koganeya, A., Nagao, K., Kurahashi, A., Kishimoto, T., Tahara, M., Yamano, A., Nagamune, K., Hirabayashi, Y., Juni, N., Umeda, M., Fujimori, F., Nishibori, K., Yamaji-Hasegawa, A., Greimel, P., Kobayashi, T., Evaluation of aegerolysins as novel tools to detect and visualize ceramide phosphoethanolamine, a major sphingolipid in invertebrates. FASEB J. 29 (9): 3920-3934 (2015). PMID 26060215. [link to RRC of NBRP]
Bhat, H.B., Kishimoto, T., Abe, M., Makino, A., Inaba, T., Murate, M., Dohmae, N., Kurahashi, A., Nishibori, K., Fujimori, F., Greimel, P., Ishitsuka, R., Kobayashi, T., Binding of a pleurotolysin ortholog from Pleurotus eryngii to sphingomyelin and cholesterol-rich membrane domains. J. Lipid Res. 54 (10): 2933-2943 (2013). PMID 23918047. [link to RRC of NBRP]

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

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 RDB13962_A6Btp1-3.pdf check

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

Primer: pBR322_BamHI_R (Pr0065)
Region: RBS, insert 5' (PlyA2, EGFP 5')
Sequence file: RDB13962_A6Btd.seq check
>D03144A5_A6Bt_1_pBR 322 BamHI R_D04_10.ab1
    1 NNNNNNNNNN NNNGNNTNNN NNTANGGNNN NTGTGAGCGG NNAACAATTC CCCTCTAGAA
   61 ATAATTTTGT TNAACTTTAA GAAGGAGATA TACCGGTACC ATGGCATACG CACAATGGGT
  121 CATCATCATC ATCCACAACG TCGGGTCCAA AGACGTGAAG ATCGTAAACC TCAAGCCTTC
  181 TTGGGGGAAG CTTCATGCCG ACGGCGACAA AGACACAGAA GTCTCAGCAA GCAAGTACGA
  241 AGGAACAGTC ATCAAGCCTG ACGAGAAGCT CCAGATCAAT GCCTGCGGCA GATCAGACGC
  301 CGCCGAAGGC ACGACAGGAA CCTTCGACCT TGTTGACCCC GCCGATGGCG ACAAGCAGGT
  361 CCGCCACTTC TACTGGGACT GCCCATGGGG AAGCAAGGCC AACACTTGGA CCGTTAGCGG
  421 TTCCAACACC AAGTGGATGA TCGAGTACTC TGGCCAGAAC TTGGACAGCG GTGCCCTCGG
  481 CACGATCACC GTCGACACCT TGAAGAAGGG GAACGAATTC GGAGGAGGAG GAGGAGGATC
  541 CATGGTGAGC AAGGGCGAGG AGCTGTTCAC CGGGGTGGTG CCCATCCTGG TCGAGCTGGA
  601 CGGCGACGTA AACGGCCACA AGTTCAGCGT GTCCGGCGAG GGCGAGGGCG ATGCCACCTA
  661 TGGCAAGCTG ANCCTGAAGT TCATCTGCAC CACCGGCAAG CTGCCCGTGC CCTGGCCCAC
  721 CCTCGTGACC ACCCTGACCT ACGGCGTGCA GTGCTTCAGC CGCTACCCCG ACCACATGAA
  781 GCAGCACGAC TTCTTCNAGT CCGCCATGCC CGAANGCTAC GTCCAGGANC GCANCATCTT
  841 CTTCNNNGAC GANNGNNACT ACAAGANCCG CGCCNANGTG NANTTCGNNG GCGANNCCCN
  901 GNNNNNNCNC ATCNAGCTGN ANNGNNNTCN NNTNNNNNNN NNANNNNNNC ATNCNGGGGN
961 ANNNGCTGGN NTNNNNCTNN NNNNNNNN
//
Primer: EGFP-N_F (Pr0649)
Region: insert 3' (EGFP 3'), 6xHis
Sequence file: RDB13962_A6Bte.seq check
>13962_13962_B2D1F_1_EGFP-N_F_A01_01_ABI08.ab1
    1 NNNNNNNNNN NGNNNNNNNG NNGGCGNNGC NNCCTATGGC AAGCTGNNCC TGAAGTTCAT
   61 CTGCACCACC GGCAAGCTGC CCGTGCCCTG GCCCACCCTC GTGACCACCC TGACCTACGG
  121 CGTGCAGTGC TTCAGCCGCT ACCCCGACCA CATGAAGCAG CACGACTTCT TCAAGTCCGC
  181 CATGCCCGAA GGCTACGTCC AGGAGCGCAC CATCTTCTTC AAGGACGACG GCAACTACAA
  241 GACCCGCGCC GAGGTGAAGT TCGAGGGCGA CACCCTGGTG AACCGCATCG AGCTGAAGGG
  301 CATCGACTTC AAGGAGGACG GCAACATCCT GGGGCACAAG CTGGAGTACA ACTACAACAG
  361 CCACAACGTC TATATCATGG CCGACAAGCA GAAGAACGGC ATCAAGGTGA ACTTCAAGAT
  421 CCGCCACAAC ATCGAGGACG GCAGCGTGCA GCTCGCCGAC CACTACCAGC AGAACACCCC
  481 CATCGGCGAC GGCCCCGTGC TGCTGCCCGA CAACCACTAC CTGAGCACCC AGTCCGCCCT
  541 GAGCAAAGAC CCCAACGAGA AGCGCGATCA CATGGTCCTG CTGGAGTTCG TGACCGCCGC
  601 CGGGATCACT CTCGGCATGG ACGAGCTGTA CAAGAAGCTT CACCACCACC ACCACCACTG
  661 AGATCCGGCT GCTAACAAAG CCCGAAAGGA AGCTGANTTG GCTGCTGCCA CCGCTGAGCA
  721 ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG AGGGGTTTTT TGCTGANAGG
  781 AGGAACTATA TCCGGANTGN CGAATGGGAC GCGCCCNGTA GCGGCGCATT AAGCGCGGCG
  841 GGTGTGGTGG TTACGCGCAG CGNGACCGCT ACACTTGCCA GCGCCCTANC GCCCGCTNCT
  901 TTCGCTTTCN NCCCTTNCTT NCNCGCNNCN TNCGCCNGCT TNCCCCGNNN NGCTCNAANN
961 CNGGGGNNNN CNNTNNNGNN NNNNNNTNNN NNNNTNNNNN NNNNN
//
Primer: EGFP-M_R (Pr0743)
Region: insert mid (EGFP 5', PlyA2 3')
Sequence file: RDB13962_A6Btf.seq check
>13962_13962_B2D1F_1_EGFP-M_R_A12_01_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNGGCGGNNC TNNNNNNNNC CTTGATGCCG TTCTTCTGCT
   61 TGTCGGCCAT GATATAGACG TTGTGGCTGT TGTAGTTGTA CTCCAGCTTG TGCCCCAGGA
  121 TGTTGCCGTC CTCCTTGAAG TCGATGCCCT TCAGCTCGAT GCGGTTCACC AGGGTGTCGC
  181 CCTCGAACTT CACCTCGGCG CGGGTCTTGT AGTTGCCGTC GTCCTTGAAG AAGATGGTGC
  241 GCTCCTGGAC GTAGCCTTCG GGCATGGCGG ACTTGAAGAA GTCGTGCTGC TTCATGTGGT
  301 CGGGGTAGCG GCTGAAGCAC TGCACGCCGT AGGTCAGGGT GGTCACGAGG GTGGGCCAGG
  361 GCACGGGCAG CTTGCCGGTG GTGCAGATGA ACTTCAGGGT CAGCTTGCCA TAGGTGGCAT
  421 CGCCCTCGCC CTCGCCGGAC ACGCTGAACT TGTGGCCGTT TACGTCGCCG TCCAGCTCGA
  481 CCAGGATGGG CACCACCCCG GTGAACAGCT CCTCGCCCTT GCTCACCATG GATCCTCCTC
  541 CTCCTCCTCC GAATTCGTTC CCCTTCTTCA AGGTGTCGAC GGTGATCGTG CCGAGGGCAC
  601 CGCTGTCCAA GTTCTGGCCA GANTACTCGA TCATCNACTT GGTGTTGGAA CCGCTNACGG
  661 TCCAAGTGNT GGNCTTGCTN CCCCNTGGGN AGTCCNNGTA NANNNGGNNN ANNNGCTNNN
721 CGNNNTCGGC GGGNNNNNNN NNN
//
Primer: pGL3_RV4 (Pr0335)
Region: lacI 3'
Sequence file: RDB13962_A6Btg.seq check
>D03144A5_A6Bt_1_pGL3_RV4_G12_21.ab1
    1 NNNNNNNNNN NNNNNNGNTC TCAGGGCNTC GGTCGANNNC 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 GTGANCCAAT GCGACCAGAT GCTCCACGCC
  661 CAGTCGCGTA CCGTCTTCAT GGGAGAAAAT AATACTGTTG ATGGGTGTCT GGTCAGANAC
  721 ATCAAGAAAT AACGCCNGAA CATTAGTGCA NGCAGCTTCC ACAGCAATGG CATCCTGGTC
  781 ATCCAGCGGA TAGTTAATGA TCAGCCCANN GANGCGTTGC GCGAGAAGNN TGTGCANCGC
  841 CGCTTTNCAG GCNTCNACGC CGCTTCNTTC TANCNTCNAC NCCACCACNC NNNNACCCAG
901 TTGNNNNNNG NNANATTTNA TCGNNNNNAN NNNTTGCNAN NNNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original Bhat, H.B., Evaluation of aegerolysins as novel tools to detect and visualize ceramide phosphoethanolamine, a major sphingolipid in invertebrates. FASEB J. 29 (9): 3920-3934 (2015). PMID 26060215. [link to RRC of NBRP]
original Bhat, H.B., Binding of a pleurotolysin ortholog from Pleurotus eryngii to sphingomyelin and cholesterol-rich membrane domains. J. Lipid Res. 54 (10): 2933-2943 (2013). PMID 23918047. [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.