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基本情報
登録情報 | ![]() | |||||||||
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タイトル | CryoEM structure of bacterial transcription intermediate complex mediated by activator PspF containing nifH promoter DNA containing mismatch from -11 to -8 - conformation 6 | |||||||||
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![]() | sigma factor / transcription / complex / AAA+ / DNA-binding protein | |||||||||
機能・相同性 | ![]() regulation of cellular response to stress / RNA polymerase complex / DNA-binding transcription activator activity / submerged biofilm formation / cellular response to cell envelope stress / regulation of DNA-templated transcription initiation / sigma factor activity / phosphorelay signal transduction system / bacterial-type flagellum assembly / bacterial-type RNA polymerase core enzyme binding ...regulation of cellular response to stress / RNA polymerase complex / DNA-binding transcription activator activity / submerged biofilm formation / cellular response to cell envelope stress / regulation of DNA-templated transcription initiation / sigma factor activity / phosphorelay signal transduction system / bacterial-type flagellum assembly / bacterial-type RNA polymerase core enzyme binding / cytosolic DNA-directed RNA polymerase complex / bacterial-type flagellum-dependent cell motility / nitrate assimilation / cis-regulatory region sequence-specific DNA binding / nucleotidyltransferase activity / DNA-directed RNA polymerase complex / transcription elongation factor complex / regulation of DNA-templated transcription elongation / transcription antitermination / DNA-templated transcription initiation / cell motility / protein-DNA complex / ribonucleoside binding / : / : / : / : / : / : / DNA-directed RNA polymerase / response to heat / protein-containing complex assembly / transcription regulator complex / sequence-specific DNA binding / intracellular iron ion homeostasis / protein dimerization activity / response to antibiotic / negative regulation of DNA-templated transcription / regulation of DNA-templated transcription / positive regulation of DNA-templated transcription / magnesium ion binding / ATP hydrolysis activity / DNA binding / zinc ion binding / ATP binding / identical protein binding / membrane / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 7.2 Å | |||||||||
![]() | Gao F / Zhang X | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Subunit specialization in AAA+ proteins and substrate unfolding during transcription complex remodeling. 著者: Forson Gao / Fuzhou Ye / Martin Buck / Xiaodong Zhang / ![]() 要旨: Bacterial RNA polymerase (RNAP) is a multisubunit enzyme that copies DNA into RNA in a process known as transcription. Bacteria use σ factors to recruit RNAP to promoter regions of genes that need ...Bacterial RNA polymerase (RNAP) is a multisubunit enzyme that copies DNA into RNA in a process known as transcription. Bacteria use σ factors to recruit RNAP to promoter regions of genes that need to be transcribed, with 60% bacteria containing at least one specialized σ factor, σ. σ recruits RNAP to promoters of genes associated with stress responses and forms a stable closed complex that does not spontaneously isomerize to the open state where promoter DNA is melted out and competent for transcription. The σ-mediated open complex formation requires specific AAA+ proteins (TPases ssociated with diverse cellular ctivities) known as bacterial enhancer-binding proteins (bEBPs). We have now obtained structures of new intermediate states of bEBP-bound complexes during transcription initiation, which elucidate the mechanism of DNA melting driven by ATPase activity of bEBPs and suggest a mechanistic model that couples the Adenosine triphosphate (ATP) hydrolysis cycle within the bEBP hexamer with σ unfolding. Our data reveal that bEBP forms a nonplanar hexamer with the hydrolysis-ready subunit located at the furthest/highest point of the spiral hexamer relative to the RNAP. ATP hydrolysis induces conformational changes in bEBP that drives a vectoral transiting of the regulatory N terminus of σ into the bEBP hexamer central pore causing the partial unfolding of σ, while forming specific bEBP contacts with promoter DNA. Furthermore, our data suggest a mechanism of the bEBP AAA+ protein that is distinct from the hand-over-hand mechanism proposed for many other AAA+ proteins, highlighting the versatile mechanisms utilized by the large protein family. | |||||||||
履歴 |
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構造の表示
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-EMDBアーカイブ
マップデータ | ![]() | 62.5 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 23.5 KB 23.5 KB | 表示 表示 | ![]() |
画像 | ![]() | 68.5 KB | ||
Filedesc metadata | ![]() | 9 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 404.3 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 403.9 KB | 表示 | |
XML形式データ | ![]() | 6.4 KB | 表示 | |
CIF形式データ | ![]() | 7.4 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 9q91MC ![]() 9q93C ![]() 9q94C ![]() 9q95C ![]() 9q96C ![]() 9q97C ![]() 9q98C C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||
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投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.1 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
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試料の構成要素
-全体 : RNA polymerase-sigma54-PspF(1-275) intermediate complex bound to ...
全体 | 名称: RNA polymerase-sigma54-PspF(1-275) intermediate complex bound to nifH DNA (-28 to +35) containing mismatch from -11 to -8, conformation 6 |
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要素 |
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-超分子 #1: RNA polymerase-sigma54-PspF(1-275) intermediate complex bound to ...
超分子 | 名称: RNA polymerase-sigma54-PspF(1-275) intermediate complex bound to nifH DNA (-28 to +35) containing mismatch from -11 to -8, conformation 6 タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#8 |
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由来(天然) | 生物種: ![]() ![]() |
-分子 #1: Psp operon transcriptional activator
分子 | 名称: Psp operon transcriptional activator / タイプ: protein_or_peptide / ID: 1 / コピー数: 6 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 29.390668 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MAEYKDNLLG EANSFLEVLE QVSHLAPLDK PVLIIGERGT GKELIASRLH YLSSRWQGPF ISLNCAALNE NLLDSELFGH EAGAFTGAQ KRHPGRFERA DGGTLFLDEL ATAPMMVQEK LLRVIEYGEL ERVGGSQPLQ VNVRLVCATN ADLPAMVNEG T FRADLLDR ...文字列: MAEYKDNLLG EANSFLEVLE QVSHLAPLDK PVLIIGERGT GKELIASRLH YLSSRWQGPF ISLNCAALNE NLLDSELFGH EAGAFTGAQ KRHPGRFERA DGGTLFLDEL ATAPMMVQEK LLRVIEYGEL ERVGGSQPLQ VNVRLVCATN ADLPAMVNEG T FRADLLDR LAFDVVQLPP LRERESDIML MAEYFAIQMC REIKLPLFPG FTERARETLL NYRWPGNIRE LKNVVERSVY RH GTSDYPL DDIIIDPFKR RP UniProtKB: Psp operon transcriptional activator |
-分子 #2: DNA-directed RNA polymerase subunit alpha
分子 | 名称: DNA-directed RNA polymerase subunit alpha / タイプ: protein_or_peptide / ID: 2 / コピー数: 2 / 光学異性体: LEVO / EC番号: DNA-directed RNA polymerase |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 36.55868 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MQGSVTEFLK PRLVDIEQVS STHAKVTLEP LERGFGHTLG NALRRILLSS MPGCAVTEVE IDGVLHEYST KEGVQEDILE ILLNLKGLA VRVQGKDEVI LTLNKSGIGP VTAADITHDG DVEIVKPQHV ICHLTDENAS ISMRIKVQRG RGYVPASTRI H SEEDERPI ...文字列: MQGSVTEFLK PRLVDIEQVS STHAKVTLEP LERGFGHTLG NALRRILLSS MPGCAVTEVE IDGVLHEYST KEGVQEDILE ILLNLKGLA VRVQGKDEVI LTLNKSGIGP VTAADITHDG DVEIVKPQHV ICHLTDENAS ISMRIKVQRG RGYVPASTRI H SEEDERPI GRLLVDACYS PVERIAYNVE AARVEQRTDL DKLVIEMETN GTIDPEEAIR RAATILAEQL EAFVDLRDVR QP EVKEEKP EFDPILLRPV DDLELTVRSA NCLKAEAIHY IGDLVQRTEV ELLKTPNLGK KSLTEIKDVL ASRGLSLGMR LEN WPPASI ADE UniProtKB: DNA-directed RNA polymerase subunit alpha |
-分子 #3: DNA-directed RNA polymerase subunit beta
分子 | 名称: DNA-directed RNA polymerase subunit beta / タイプ: protein_or_peptide / ID: 3 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 150.69175 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MVYSYTEKKR IRKDFGKRPQ VLDVPYLLSI QLDSFQKFIE QDPEGQYGLE AAFRSVFPIQ SYSGNSELQY VSYRLGEPVF DVQECQIRG VTYSAPLRVK LRLVIYEREA PEGTVKDIKE QEVYMGEIPL MTDNGTFVIN GTERVIVSQL HRSPGVFFDS D KGKTHSSG ...文字列: MVYSYTEKKR IRKDFGKRPQ VLDVPYLLSI QLDSFQKFIE QDPEGQYGLE AAFRSVFPIQ SYSGNSELQY VSYRLGEPVF DVQECQIRG VTYSAPLRVK LRLVIYEREA PEGTVKDIKE QEVYMGEIPL MTDNGTFVIN GTERVIVSQL HRSPGVFFDS D KGKTHSSG KVLYNARIIP YRGSWLDFEF DPKDNLFVRI DRRRKLPATI ILRALNYTTE QILDLFFEKV IFEIRDNKLQ ME LVPERLR GETASFDIEA NGKVYVEKGR RITARHIRQL EKDDVKLIEV PVEYIAGKVV AKDYIDESTG ELICAANMEL SLD LLAKLS QSGHKRIETL FTNDLDHGPY ISETLRVDPT NDRLSALVEI YRMMRPGEPP TREAAESLFE NLFFSEDRYD LSAV GRMKF NRSLLREEIE GSGILSKDDI IDVMKKLIDI RNGKGEVDDI DHLGNRRIRS VGEMAENQFR VGLVRVERAV KERLS LGDL DTLMPQDMIN AKPISAAVKE FFGSSQLSQF MDQNNPLSEI THKRRISALG PGGLTRERAG FEVRDVHPTH YGRVCP IET PEGPNIGLIN SLSVYAQTNE YGFLETPYRK VTDGVVTDEI HYLSAIEEGN YVIAQANSNL DEEGHFVEDL VTCRSKG ES SLFSRDQVDY MDVSTQQVVS VGASLIPFLE HDDANRALMG ANMQRQAVPT LRADKPLVGT GMERAVAVDS GVTAVAKR G GVVQYVDASR IVIKVNEDEM YPGEAGIDIY NLTKYTRSNQ NTCINQMPCV SLGEPVERGD VLADGPSTDL GELALGQNM RVAFMPWNGY NFEDSILVSE RVVQEDRFTT IHIQELACVS RDTKLGPEEI TADIPNVGEA ALSKLDESGI VYIGAEVTGG DILVGKVTP KGETQLTPEE KLLRAIFGEK ASDVKDSSLR VPNGVSGTVI DVQVFTRDGV EKDKRALEIE EMQLKQAKKD L SEELQILE AGLFSRIRAV LVAGGVEAEK LDKLPRDRWL ELGLTDEEKQ NQLEQLAEQY DELKHEFEKK LEAKRRKITQ GD DLAPGVL KIVKVYLAVK RRIQPGDKMA GRHGNKGVIS KINPIEDMPY DENGTPVDIV LNPLGVPSRM NIGQILETHL GMA AKGIGD KINAMLKQQQ EVAKLREFIQ RAYDLGADVR QKVDLSTFSD EEVMRLAENL RKGMPIATPV FDGAKEAEIK ELLK LGDLP TSGQIRLYDG RTGEQFERPV TVGYMYMLKL NHLVDDKMHA RSTGSYSLVT QQPLGGKAQF GGQRFGEMEV WALEA YGAA YTLQEMLTVK SDDVNGRTKM YKNIVDGNHQ MEPGMPESFN VLLKEIRSLG INIELED UniProtKB: DNA-directed RNA polymerase subunit beta |
-分子 #4: DNA-directed RNA polymerase subunit omega
分子 | 名称: DNA-directed RNA polymerase subunit omega / タイプ: protein_or_peptide / ID: 4 / コピー数: 1 / 光学異性体: LEVO / EC番号: DNA-directed RNA polymerase |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 10.249547 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MARVTVQDAV EKIGNRFDLV LVAARRARQM QVGGKDPLVP EENDKTTVIA LREIEEGLIN NQILDVRERQ EQQEQEAAEL QAVTAIAEG RR UniProtKB: DNA-directed RNA polymerase subunit omega |
-分子 #7: RNA polymerase sigma-54 factor
分子 | 名称: RNA polymerase sigma-54 factor / タイプ: protein_or_peptide / ID: 7 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 53.722402 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MKQGLQLRLS LAMTPQLQQA IRLLQLSTLE LQQELQQALE SNPLLEQTDL HDEVEAKEVE DRESLDTVDA LEQKEMPDEL PLDASWDEI YTAGTPSGNG VDYQDDELPV YQGETTQTLQ DYLMWQVELT PFTDTDRAIA TSIVDAVDDT GYLTIQIEDI V DSIGDDEI ...文字列: MKQGLQLRLS LAMTPQLQQA IRLLQLSTLE LQQELQQALE SNPLLEQTDL HDEVEAKEVE DRESLDTVDA LEQKEMPDEL PLDASWDEI YTAGTPSGNG VDYQDDELPV YQGETTQTLQ DYLMWQVELT PFTDTDRAIA TSIVDAVDDT GYLTIQIEDI V DSIGDDEI GLEEVEAVLK RIQRFDPVGV AAKDLRDCLL IQLSQFAKET PWLEEARLII SDHLDLLANH DFRTLMRVTR LK EEVLKEA VNLIQSLDPR PGQSIQTSEP EYVIPDVLVR KVSGRWTVEL NADSIPRLKI NQQYAAMGNS ARNDADGQFI RSN LQEARW LIKSLESRND TLLRVSRCIV EQQQAFFEQG EEYMKPMVLA DIAQAVEMHE STISRVTTQK YLHSPRGIFE LKYF FSSHV NTEGGGEASS TAIRALVKKL IAAENPAKPL SDSKLTSMLS EQGIMVARRT VAKYRESLSI PPSNQRKQLV UniProtKB: RNA polymerase sigma-54 factor |
-分子 #8: DNA-directed RNA polymerase subunit beta'
分子 | 名称: DNA-directed RNA polymerase subunit beta' / タイプ: protein_or_peptide / ID: 8 / コピー数: 1 / 光学異性体: LEVO / EC番号: DNA-directed RNA polymerase |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 155.366781 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MKDLLKFLKA QTKTEEFDAI KIALASPDMI RSWSFGEVKK PETINYRTFK PERDGLFCAR IFGPVKDYEC LCGKYKRLKH RGVICEKCG VEVTQTKVRR ERMGHIELAS PTAHIWFLKS LPSRIGLLLD MPLRDIERVL YFESYVVIEG GMTNLERQQI L TEEQYLDA ...文字列: MKDLLKFLKA QTKTEEFDAI KIALASPDMI RSWSFGEVKK PETINYRTFK PERDGLFCAR IFGPVKDYEC LCGKYKRLKH RGVICEKCG VEVTQTKVRR ERMGHIELAS PTAHIWFLKS LPSRIGLLLD MPLRDIERVL YFESYVVIEG GMTNLERQQI L TEEQYLDA LEEFGDEFDA KMGAEAIQAL LKSMDLEQEC EQLREELNET NSETKRKKLT KRIKLLEAFV QSGNKPEWMI LT VLPVLPP DLRPLVPLDG GRFATSDLND LYRRVINRNN RLKRLLDLAA PDIIVRNEKR MLQEAVDALL DNGRRGRAIT GSN KRPLKS LADMIKGKQG RFRQNLLGKR VDYSGRSVIT VGPYLRLHQC GLPKKMALEL FKPFIYGKLE LRGLATTIKA AKKM VEREE AVVWDILDEV IREHPVLLNR APTLHRLGIQ AFEPVLIEGK AIQLHPLVCA AYNADFDGDQ MAVHVPLTLE AQLEA RALM MSTNNILSPA NGEPIIVPSQ DVVLGLYYMT RDCVNAKGEG MVLTGPKEAE RLYRSGLASL HARVKVRITE YEKDAN GEL VAKTSLKDTT VGRAILWMIV PKGLPYSIVN QALGKKAISK MLNTCYRILG LKPTVIFADQ IMYTGFAYAA RSGASVG ID DMVIPEKKHE IISEAEAEVA EIQEQFQSGL VTAGERYNKV IDIWAAANDR VSKAMMDNLQ TETVINRDGQ EEKQVSFN S IYMMADSGAR GSAAQIRQLA GMRGLMAKPD GSIIETPITA NFREGLNVLQ YFISTHGARK GLADTALKTA NSGYLTRRL VDVAQDLVVT EDDCGTHEGI MMTPVIEGGD VKEPLRDRVL GRVTAEDVLK PGTADILVPR NTLLHEQWCD LLEENSVDAV KVRSVVSCD TDFGVCAHCY GRDLARGHII NKGEAIGVIA AQSIGEPGTQ LTMRTFHIGG AASRAAAESS IQVKNKGSIK L SNVKSVVN SSGKLVITSR NTELKLIDEF GRTKESYKVP YGAVLAKGDG EQVAGGETVA NWDPHTMPVI TEVSGFVRFT DM IDGQTIT RQTDELTGLS SLVVLDSAER TAGGKDLRPA LKIVDAQGND VLIPGTDMPA QYFLPGKAIV QLEDGVQISS GDT LARIPQ ESGGTKDITG GLPRVADLFE ARRPKEPAIL AEISGIVSFG KETKGKRRLV ITPVDGSDPY EEMIPKWRQL NVFE GERVE RGDVISDGPE APHDILRLRG VHAVTRYIVN EVQDVYRLQG VKINDKHIEV IVRQMLRKAT IVNAGSSDFL EGEQV EYSR VKIANRELEA NGKVGATYSR DLLGITKASL ATESFISAAS FQETTRVLTE AAVAGKRDEL RGLKENVIVG RLIPAG TGY AYHQDRMRRR AAGEAPAAPQ VTAEDASASL AELLNAGLGG SDNE UniProtKB: DNA-directed RNA polymerase subunit beta' |
-分子 #5: Non-template DNA (34-MER)
分子 | 名称: Non-template DNA (34-MER) / タイプ: dna / ID: 5 / コピー数: 1 / 分類: DNA |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 10.420691 KDa |
配列 | 文字列: (DA)(DG)(DA)(DC)(DG)(DG)(DC)(DT)(DG)(DG) (DC)(DA)(DC)(DG)(DA)(DC)(DT)(DT)(DT)(DT) (DG)(DC)(DA)(DA)(DT)(DC)(DG)(DC)(DA) (DG)(DC)(DC)(DC)(DT) |
-分子 #6: Template DNA (34-MER)
分子 | 名称: Template DNA (34-MER) / タイプ: dna / ID: 6 / コピー数: 1 / 分類: DNA |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 10.50074 KDa |
配列 | 文字列: (DA)(DG)(DG)(DG)(DC)(DT)(DG)(DA)(DT)(DC) (DG)(DT)(DG)(DC)(DA)(DA)(DA)(DA)(DG)(DT) (DC)(DG)(DT)(DG)(DC)(DC)(DA)(DG)(DC) (DC)(DG)(DT)(DC)(DT) |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 8 |
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凍結 | 凍結剤: ETHANE |
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電子顕微鏡法
顕微鏡 | TFS KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 平均電子線量: 50.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 3.0 µm / 最小 デフォーカス(公称値): 1.0 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |