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rdf:type
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Description
| - To obtain strong bacterial expression of proteins that seem to be hard to express in bacteria or are highly toxic for bacteria, it is possible to vreate a palette of similar constructs, differing only by several nucleotides, gradually deleted from the full-lenght clone by ewonuclease III. When a construct is equipped with the 6xHis tag, a simple colony-blot procedure can be performed and a colony giving strong and efficient expression can easily sbe selected for high range protein expression. We utilized this procedure to produce one of potato mop-top virus (PMTV) movement proteins, namely triple gene block protein 1 (TGPp1) which was very hard toexpress in bacteria in its original length. The TGPp1 gene was digested with exonuclease III and nuclease S1 from its 5´ terminus. Leaving 6xHis tag intact. The clone that showed the strongest signal with anti-His antibodies in colony-blot procedure was found to have 44 amino acids (of total 463) deleted. The SDS-PAGE and Western blot of h...
- To obtain strong bacterial expression of proteins that seem to be hard to express in bacteria or are highly toxic for bacteria, it is possible to vreate a palette of similar constructs, differing only by several nucleotides, gradually deleted from the full-lenght clone by ewonuclease III. When a construct is equipped with the 6xHis tag, a simple colony-blot procedure can be performed and a colony giving strong and efficient expression can easily sbe selected for high range protein expression. We utilized this procedure to produce one of potato mop-top virus (PMTV) movement proteins, namely triple gene block protein 1 (TGPp1) which was very hard toexpress in bacteria in its original length. The TGPp1 gene was digested with exonuclease III and nuclease S1 from its 5´ terminus. Leaving 6xHis tag intact. The clone that showed the strongest signal with anti-His antibodies in colony-blot procedure was found to have 44 amino acids (of total 463) deleted. The SDS-PAGE and Western blot of h... (en)
- Byla vypracována metoda pro expresi těžko exprimovatelných či toxických proteinů v bakteriích. Pomocí postupného zkracování exonukleasou III a nukleasou S1 z 5´konce se vytvořila škála podobných konstruktů lišících se pouze o několik nukleotidů. Konstrukty byly vybaveny 6 x His kotvou, která umožnila jejich detekci v colony- blotu. Pro expresi ve velkém měřítku se poté vybraly kolonie poskytující dostatečně silnou expresi. Tato metoda byla využita pro přípravu jednoho z nestrukturních „movement“ proteinů mop-top viru bramboru (PMTV) ( prvního proteinu z „triple gene block“ (TGBp1)).Klon, který vykazoval nejsilnější expresi byl zkrácen o 44 aminokyselin z celkových 463. (cs)
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Title
| - Efficient bacterial expression of recombinant potato mop-top virus non-structural triple gene block protein 1 modified by progressive deletion of its N-terminus
- Efficient bacterial expression of recombinant potato mop-top virus non-structural triple gene block protein 1 modified by progressive deletion of its N-terminus (en)
- Účinná bakteriální exprese rekombinantního nestrukturálního proteinu 1 z „triple gene blocku“ mop-top viru bramboru (cs)
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skos:prefLabel
| - Efficient bacterial expression of recombinant potato mop-top virus non-structural triple gene block protein 1 modified by progressive deletion of its N-terminus
- Efficient bacterial expression of recombinant potato mop-top virus non-structural triple gene block protein 1 modified by progressive deletion of its N-terminus (en)
- Účinná bakteriální exprese rekombinantního nestrukturálního proteinu 1 z „triple gene blocku“ mop-top viru bramboru (cs)
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skos:notation
| - RIV/61389030:_____/05:00028432!RIV06-GA0-61389030
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http://linked.open.../vavai/riv/strany
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http://linked.open...avai/riv/aktivita
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http://linked.open...avai/riv/aktivity
| - P(GA522/04/1329), Z(AV0Z50380511)
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http://linked.open...iv/cisloPeriodika
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http://linked.open...vai/riv/dodaniDat
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http://linked.open...aciTvurceVysledku
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http://linked.open.../riv/druhVysledku
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http://linked.open...iv/duvernostUdaju
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http://linked.open...titaPredkladatele
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http://linked.open...dnocenehoVysledku
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http://linked.open...ai/riv/idVysledku
| - RIV/61389030:_____/05:00028432
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http://linked.open...riv/jazykVysledku
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http://linked.open.../riv/klicovaSlova
| - Protein expression; Potato mop-top virus; Triple gene block (en)
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http://linked.open.../riv/klicoveSlovo
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http://linked.open...odStatuVydavatele
| - US - Spojené státy americké
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http://linked.open...ontrolniKodProRIV
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http://linked.open...i/riv/nazevZdroje
| - Protein Expression and Purification
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http://linked.open...in/vavai/riv/obor
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http://linked.open...ichTvurcuVysledku
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http://linked.open...cetTvurcuVysledku
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http://linked.open...vavai/riv/projekt
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http://linked.open...UplatneniVysledku
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http://linked.open...v/svazekPeriodika
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http://linked.open...iv/tvurceVysledku
| - Čeřovská, Noemi
- Pečenková, Tamara
- Filigarová, Marie
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http://linked.open...n/vavai/riv/zamer
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issn
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number of pages
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is http://linked.open...avai/riv/vysledek
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