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Yorodumi- EMDB-54012: Structure of the Bacteriophage PhiKZ non-virion RNA Polymerase bo... -
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Open data
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Basic information
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| Title | Structure of the Bacteriophage PhiKZ non-virion RNA Polymerase bound to an analogue of its promoter | |||||||||
Map data | PhiKZ nvRNAP bound to promoter analogue - RELION gold-standard map | |||||||||
Sample |
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Keywords | PhiKZ / nvRNAP / RNA / DNA / transcription / RNA BINDING PROTEIN | |||||||||
| Function / homology | DNA-directed RNA polymerase / PHIKZ056.1 / PHIKZ123 / PHIKZ074 / PHIKZ068 Function and homology information | |||||||||
| Biological species | Pseudomonas phage phiKZ (virus) / Phikzvirus phiKZ | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.59 Å | |||||||||
Authors | Chen C / de Martin Garrido N / Yakunina M / Aylett CHS | |||||||||
| Funding support | United Kingdom, 2 items
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Citation | Journal: J Mol Biol / Year: 2024Title: Structure of the Bacteriophage PhiKZ Non-virion RNA Polymerase Transcribing from its Promoter p119L. Authors: Natàlia de Martín Garrido / Chao-Sheng Chen / Kailash Ramlaul / Christopher H S Aylett / Maria Yakunina / ![]() Abstract: Bacteriophage ΦKZ (PhiKZ) is the founding member of a family of giant bacterial viruses. It has potential as a therapeutic as its host, Pseudomonas aeruginosa, kills tens of thousands of people ...Bacteriophage ΦKZ (PhiKZ) is the founding member of a family of giant bacterial viruses. It has potential as a therapeutic as its host, Pseudomonas aeruginosa, kills tens of thousands of people worldwide each year. ΦKZ infection is independent of the host transcriptional apparatus; the virus forms a "nucleus", producing a proteinaceous barrier around the ΦKZ genome that excludes the host immune systems. It expresses its own non-canonical multi-subunit non-virion RNA polymerase (nvRNAP), which is imported into its "nucleus" to transcribe viral genes. The ΦKZ nvRNAP is formed by four polypeptides representing homologues of the eubacterial β/β' subunits, and a fifth that is likely to have evolved from an ancestral homologue to σ-factor. We have resolved the structure of the ΦKZ nvRNAP initiating transcription from its cognate promoter, p119L, including previously disordered regions. Our results shed light on the similarities and differences between ΦKZ nvRNAP mechanisms of transcription and those of canonical eubacterial RNAPs and the related non-canonical nvRNAP of bacteriophage AR9. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_54012.map.gz | 71.2 MB | EMDB map data format | |
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| Header (meta data) | emd-54012-v30.xml emd-54012.xml | 33.5 KB 33.5 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_54012_fsc.xml | 10.2 KB | Display | FSC data file |
| Images | emd_54012.png | 120.4 KB | ||
| Masks | emd_54012_msk_1.map | 91.1 MB | Mask map | |
| Filedesc metadata | emd-54012.cif.gz | 9.7 KB | ||
| Others | emd_54012_additional_1.map.gz emd_54012_half_map_1.map.gz emd_54012_half_map_2.map.gz | 85.3 MB 71.3 MB 71.3 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-54012 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-54012 | HTTPS FTP |
-Validation report
| Summary document | emd_54012_validation.pdf.gz | 922.6 KB | Display | EMDB validaton report |
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| Full document | emd_54012_full_validation.pdf.gz | 922.1 KB | Display | |
| Data in XML | emd_54012_validation.xml.gz | 17.4 KB | Display | |
| Data in CIF | emd_54012_validation.cif.gz | 22.9 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-54012 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-54012 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9rjsMC ![]() 8queC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_54012.map.gz / Format: CCP4 / Size: 91.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | PhiKZ nvRNAP bound to promoter analogue - RELION gold-standard map | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.99 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Mask #1
| File | emd_54012_msk_1.map | ||||||||||||
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-Additional map: PhiKZ nvRNAP bound to promoter analogue - sharpened 2.5A map
| File | emd_54012_additional_1.map | ||||||||||||
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| Annotation | PhiKZ nvRNAP bound to promoter analogue - sharpened 2.5A map | ||||||||||||
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| Density Histograms |
-Half map: PhiKZ nvRNAP bound to promoter analogue - RELION...
| File | emd_54012_half_map_1.map | ||||||||||||
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| Annotation | PhiKZ nvRNAP bound to promoter analogue - RELION gold-standard half map 2 | ||||||||||||
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| Density Histograms |
-Half map: PhiKZ nvRNAP bound to promoter analogue - RELION...
| File | emd_54012_half_map_2.map | ||||||||||||
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| Annotation | PhiKZ nvRNAP bound to promoter analogue - RELION gold-standard half map 1 | ||||||||||||
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| Density Histograms |
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Sample components
-Entire : PhiKZ non-virion RNA polymerase bound to an analogue of promoter p119L
| Entire | Name: PhiKZ non-virion RNA polymerase bound to an analogue of promoter p119L |
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| Components |
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-Supramolecule #1: PhiKZ non-virion RNA polymerase bound to an analogue of promoter p119L
| Supramolecule | Name: PhiKZ non-virion RNA polymerase bound to an analogue of promoter p119L type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#7 Details: Mismatches introduced around the transcription start site to stably open the bubble |
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| Source (natural) | Organism: Pseudomonas phage phiKZ (virus) |
| Molecular weight | Theoretical: 400 KDa |
-Macromolecule #1: DNA-directed RNA polymerase,PHIKZ056.1
| Macromolecule | Name: DNA-directed RNA polymerase,PHIKZ056.1 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Phikzvirus phiKZ |
| Molecular weight | Theoretical: 57.976605 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MGSSHHHHHH SSGLVPRGSH MGLYAKVVDH NEVHDQFTGK RIYANDYNTS NSDEKEEFDR HFYSHFQDSE AIESSVSCDC RAIEDAHKL GVICDICNTP VVNTSSRPIE PSMWVRTPKH VRSLINPRLI IMLTGYLVTK EFDFLAYLTD TSYRYDVESI G SKETRRKV ...String: MGSSHHHHHH SSGLVPRGSH MGLYAKVVDH NEVHDQFTGK RIYANDYNTS NSDEKEEFDR HFYSHFQDSE AIESSVSCDC RAIEDAHKL GVICDICNTP VVNTSSRPIE PSMWVRTPKH VRSLINPRLI IMLTGYLVTK EFDFLAYLTD TSYRYDVESI G SKETRRKV DRLLHRGFER GLNHFIDNFN EIFQFLLDAN IISNNKSEFA QFVAQNKDKL FPKYLPVPSK LCFVAESTTS GT YLDKPIE AAIDATLTFA SIDASSVPLS PIKAQNRTMR GLRLYGQFYE IYAKSRIAQK PGLARRHMFG ARLNATARAV ITS ISDPHD YDELHIPWGV GCQLLKYHLT NKLKAKFNMT TREAFSFVYE NVLQYNQIIA DLFKELIAEA APYKGMGCTF HRNP TLQRG STQQFFITKV KDDINDNSIS MSVLCLKAPN ADFDGDQLNL TLMPDVYLTK ATERIAPHTW VLSIDEPHEI SGNLE LQGP VVETIINWAH EKYLPPLEEW LKAK UniProtKB: DNA-directed RNA polymerase, PHIKZ056.1 |
-Macromolecule #2: PHIKZ068
| Macromolecule | Name: PHIKZ068 / type: protein_or_peptide / ID: 2 / Details: Numbering correct in original mmCIF file / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Phikzvirus phiKZ |
| Molecular weight | Theoretical: 59.442824 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MEIIVTGVQG TGFTEVATEH NGKRLTWTTT AYSKIRVQDQ QRVFQEINDY WSGLSAEAQQ HIWNCYVEIR KIMDMAMHPM RIAMSLSYY IKEMYKAMPM NSFRRWLLTI GKLYIPVDIE EVITDDSRYN RPDQTYLKHD YINLASVSLA LRPLVPIWGE F IDQGTSQE ...String: MEIIVTGVQG TGFTEVATEH NGKRLTWTTT AYSKIRVQDQ QRVFQEINDY WSGLSAEAQQ HIWNCYVEIR KIMDMAMHPM RIAMSLSYY IKEMYKAMPM NSFRRWLLTI GKLYIPVDIE EVITDDSRYN RPDQTYLKHD YINLASVSLA LRPLVPIWGE F IDQGTSQE MHKECEVISL ISDCEVNHWP VDEISIDGTP VETAYDKLSA YVKFCVEDEA PTLANLYRGM SSAEVPDILQ AK VMVRRLT ILPLNDATSH SIVSNMFRYV KSNLNPAERS TADRVNDKRP DKGGIDDDDK TSFIESHKTK QRVTPGDIVA YNL DALDVV KLVHKIDDTV PVELIQECLD CVAVTATKDI YPHQILLAQW VMHKAFPARA FSHINKNAVN HLLAAAQSLM WHWG FQQVA VFMQVELYYS GEHAMSIQPR NSTRIQIKYK DVMDELYPHQ RQQRAINGVP VAPVNIAGIA VQSAHASIRS SNWIY HGPD RLFKEAEQVT QNKVLVVPAT IKSVITELVI HLGKLNQ UniProtKB: PHIKZ068 |
-Macromolecule #3: PHIKZ071
| Macromolecule | Name: PHIKZ071 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Phikzvirus phiKZ |
| Molecular weight | Theoretical: 78.780453 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MSQLGRREID LTLLGHTGLD PWYGTTSSAR GAMFVTHIGQ APEVNGNESR YFLTGAELEY AKYTHDVRFP EDCRVLHVLR KYPTGIGKD SIRSNPVTTI IYENYFDKYK TIGVLHVPEY MSHHQDFGYE LVKNREVWET IAPNEMFSKD TVIAQSGAVK K DGTLGMGV ...String: MSQLGRREID LTLLGHTGLD PWYGTTSSAR GAMFVTHIGQ APEVNGNESR YFLTGAELEY AKYTHDVRFP EDCRVLHVLR KYPTGIGKD SIRSNPVTTI IYENYFDKYK TIGVLHVPEY MSHHQDFGYE LVKNREVWET IAPNEMFSKD TVIAQSGAVK K DGTLGMGV NANVVFLSAA GTIEDGFVAN KNFLKRMMPT SYSTAVANAG RKAFFLNMYG DDKIYKPFPD IGDVIRPDGV IF AIRDHDD DLAPAEMTPR ALRTLDRTFD RAVIGTPGAK VIDIDIWRDE RVNPSPTPTG MDAQLVKYHT HLSSYYRELL KIY RGLLAR RKDDLHITEE FERLIVTAQM FLPQPDNVRK LSRFYRLDPL DEWRVEVTYK AQKMPAGAFK MTDFHGGKGV ICKV MEDED MPIDENGNRA DLIIFGGSTM RRSNYGRIYE HGFGAAARDL AQRLRVEAGL DRHAKPTQQQ LNSVMGNTQW VDYAF KELL GFYEIIAPTM HSKMMEHPNP AEHVKTVLMD GFPYIYAPVD DPVDLMAAVN KLINSDKYRP HYGKVSYRDQ AGKWVT TKD NVLMGPLYMM LLEKIGEDWS AAASVKTQPF GLPSKLNNAD RASTPGRETA IRSFGESETR SYNCTVGPGP TAEILDQ TN NPLAHAAVIE SWLTAEKPSS VPVAVDREKI PFGGSRPVAM FDHLLECSGI ALEYAPDH |
-Macromolecule #4: PHIKZ074
| Macromolecule | Name: PHIKZ074 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Phikzvirus phiKZ |
| Molecular weight | Theoretical: 77.513461 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MNLNRYKARD LLNLSYDDLW SLPSEWHLIE FDDGKTVVSV DRITKLSVLC WYPLKHYKDC PIPSDHHIDF NRILTDNPKD YLNVEGGRV TSKAMVKHLN KAIWNIYDWS GETVDPEVLS KLAIEGKNWL YNQTTVKLSE YLATLSMFDI AEVYNHPKVR E ANHNIEPT ...String: MNLNRYKARD LLNLSYDDLW SLPSEWHLIE FDDGKTVVSV DRITKLSVLC WYPLKHYKDC PIPSDHHIDF NRILTDNPKD YLNVEGGRV TSKAMVKHLN KAIWNIYDWS GETVDPEVLS KLAIEGKNWL YNQTTVKLSE YLATLSMFDI AEVYNHPKVR E ANHNIEPT TYGIEKISYG KVKEVFNDPT QFIGNSIIEG LRSGTQKTEQ LLQAFAWRGF PTDINSDIFK YPVTTGYIDG IW NLYENMI ESRSGTKALL YNKELLRVTE YFNRKSQLIA QYVQRLHPGD CKTTILAEYP VTKLTLKAFK GKYYQKEDGK LDW IRGNET HLIGTKQKFR SVFGCNHPDS QGICMTCYGR LGINIPKGTN IGQVAAVSMG DKITSAVLST KHTDASSAVE QYKL GKIES NYLRTGEIPE TLYLKKELTQ KDYRLVIARS EAENLADILM IDDLTAYPAT SATELTSLAL VYDDEVNGEC GDVLT VSLY NRRASLSIEM LKHIKMVRWE LDQRDNIVIS LRGFDFNLPF LTLPNKHVNM YEVMKRFQSF LHSGSDSAEA GKLSTE KVG YTSKTYLKNY KSPIEALPVF ATMANEKISL NISHCEILIY AMMIRSAQYR DYRLPKPGIN GQFEKYNRLM QCRSLGG AM AFEKQHEPLN NPGSFLNKMR NDHPYDLLVK GGKLR UniProtKB: PHIKZ074 |
-Macromolecule #5: PHIKZ123
| Macromolecule | Name: PHIKZ123 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Phikzvirus phiKZ |
| Molecular weight | Theoretical: 62.95909 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MPDPFLIEKI RENTPCMNPT LANGITVEHT MTRDPNTGVN MTRRYIDSLF DISSVLFPDG FKYEGNRACT PLKHFEEITR EYNAKRIAN IAPTDMYMID LMFSYKGEML YPRPMLLPAF KRGNMVTING AKYIGSPVLT DVGFSVLNDS IFIPFRRTKL T FKQTDHHY ...String: MPDPFLIEKI RENTPCMNPT LANGITVEHT MTRDPNTGVN MTRRYIDSLF DISSVLFPDG FKYEGNRACT PLKHFEEITR EYNAKRIAN IAPTDMYMID LMFSYKGEML YPRPMLLPAF KRGNMVTING AKYIGSPVLT DVGFSVLNDS IFIPFRRTKL T FKQTDHHY MCNGQRKIMY VIWSQIHNEM AKRTKRDLGN RPHIESCLAH YFFCQFGVTQ TFKQWANVDV KCGLLSDFPE EE YPREKWN IYSSATLKGK HPTGEMVLVI PRHQESIFAT RLIAGFWYVV DAFPMRFTRP EYVDSTNLWR VILGHMVFGD FEH QGKVEE NIDSHLHSFC NSLDEMTIEE LKTVGVNVST IWELLYEIMT SLAHHLYATD IDETSMYGKR LTVLHYLMSE FNYA VSMFG YMFQSRRDRE WTVQELNEGL KRSFKLQTAI KRLTVDHGEL DTMSNPNSSM LIKGTSILVT QDRAKTAKAH NKSLI NDSS RIIHASIAEV GQYKNQPKNN PDGRGRLNMY TKVGPTGLVE RREEVREIID NAQLMFRAK UniProtKB: PHIKZ123 |
-Macromolecule #6: DNA - ATGAGTAATTTTAGTGAATGTATTTGCTATATTGCTATGTAGACAGTTCCCAAAAGCCT...
| Macromolecule | Name: DNA - ATGAGTAATTTTAGTGAATGTATTTGCTATATTGCTATGTAGACAGTTCCCAAAAGCCTAAAGTTACAATATAGG type: dna / ID: 6 / Number of copies: 1 / Classification: DNA |
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| Source (natural) | Organism: Phikzvirus phiKZ |
| Molecular weight | Theoretical: 23.218951 KDa |
| Sequence | String: (DA)(DT)(DG)(DA)(DG)(DT)(DA)(DA)(DT)(DT) (DT)(DT)(DA)(DG)(DT)(DG)(DA)(DA)(DT)(DG) (DT)(DA)(DT)(DT)(DT)(DG)(DC)(DT)(DA) (DT)(DA)(DT)(DT)(DG)(DC)(DT)(DA)(DT)(DG) (DT) (DA)(DG)(DA)(DC)(DA)(DG) ...String: (DA)(DT)(DG)(DA)(DG)(DT)(DA)(DA)(DT)(DT) (DT)(DT)(DA)(DG)(DT)(DG)(DA)(DA)(DT)(DG) (DT)(DA)(DT)(DT)(DT)(DG)(DC)(DT)(DA) (DT)(DA)(DT)(DT)(DG)(DC)(DT)(DA)(DT)(DG) (DT) (DA)(DG)(DA)(DC)(DA)(DG)(DT)(DT) (DC)(DC)(DC)(DA)(DA)(DA)(DA)(DG)(DC)(DC) (DT)(DA) (DA)(DA)(DG)(DT)(DT)(DA)(DC) (DA)(DA)(DT)(DA)(DT)(DA)(DG)(DG) |
-Macromolecule #7: DNA - CCTATATTGTAACTTTAGGCTTTTGGGAACTCCTCTCATATTCCCATAGCAAATACATT...
| Macromolecule | Name: DNA - CCTATATTGTAACTTTAGGCTTTTGGGAACTCCTCTCATATTCCCATAGCAAATACATTCACTAAAATTACTCAT type: dna / ID: 7 / Number of copies: 1 / Classification: DNA |
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| Source (natural) | Organism: Phikzvirus phiKZ |
| Molecular weight | Theoretical: 22.881719 KDa |
| Sequence | String: (DC)(DC)(DT)(DA)(DT)(DA)(DT)(DT)(DG)(DT) (DA)(DA)(DC)(DT)(DT)(DT)(DA)(DG)(DG)(DC) (DT)(DT)(DT)(DT)(DG)(DG)(DG)(DA)(DA) (DC)(DT)(DC)(DC)(DT)(DC)(DT)(DC)(DA)(DT) (DA) (DT)(DT)(DC)(DC)(DC)(DA) ...String: (DC)(DC)(DT)(DA)(DT)(DA)(DT)(DT)(DG)(DT) (DA)(DA)(DC)(DT)(DT)(DT)(DA)(DG)(DG)(DC) (DT)(DT)(DT)(DT)(DG)(DG)(DG)(DA)(DA) (DC)(DT)(DC)(DC)(DT)(DC)(DT)(DC)(DA)(DT) (DA) (DT)(DT)(DC)(DC)(DC)(DA)(DT)(DA) (DG)(DC)(DA)(DA)(DA)(DT)(DA)(DC)(DA)(DT) (DT)(DC) (DA)(DC)(DT)(DA)(DA)(DA)(DA) (DT)(DT)(DA)(DC)(DT)(DC)(DA)(DT) |
-Macromolecule #8: ZINC ION
| Macromolecule | Name: ZINC ION / type: ligand / ID: 8 / Number of copies: 2 / Formula: ZN |
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| Molecular weight | Theoretical: 65.409 Da |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.1 mg/mL | ||||||||||||
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| Buffer | pH: 8 Component:
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| Grid | Model: Quantifoil R2/1 / Material: GOLD / Mesh: 300 / Support film - Material: GRAPHENE OXIDE / Support film - topology: CONTINUOUS / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR | ||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 289 K / Instrument: FEI VITROBOT MARK IV Details: Settings: -4 blot force, 4 s waiting time and 0.5-1 s blotting time.. | ||||||||||||
| Details | PhiKZ nvRNAP complexes were mixed at 0.1 mg/mL final concentration with the P119L promoter analogue at a 1:1 ratio. Reactions were incubated for 30 min at 25 C before use. |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 10 eV |
| Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number grids imaged: 1 / Number real images: 20670 / Average electron dose: 50.0 e/Å2 / Details: Images collected as TIFF movies |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.75 µm / Nominal magnification: 130000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model |
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| Refinement | Space: REAL / Protocol: AB INITIO MODEL / Overall B value: 104.82 / Target criteria: CC | ||||||||||||||||||
| Output model | ![]() PDB-9rjs: |
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About Yorodumi



Keywords
Pseudomonas phage phiKZ (virus)
Authors
United Kingdom, 2 items
Citation



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FIELD EMISSION GUN



