Screening mutant libraries of T7 RNA polymerase for candidates with increased acceptance of 2'-modified nueleotidest

Vanessa Siegmund; Tobias Santner; Ronald Micura; Andreas Marx

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Screening mutant libraries of T7 RNA polymerase for candidates with increased acceptance of 2'-modified nueleotidest

机译：筛选T7 RNA聚合酶突变文库中2'-修饰的核糖体接受度增加的候选对象

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We present a screening assay based on fluorescence readout for the directed evolution of T7 RNA polymerase variants with acceptance of 2-modified nucleotides. By using this screening we were able to identify a T7 RNA polymerase mutant with increased acceptance of 2'-methylseleno-2'-deoxyuridine 5'-triphosphate.

机译：我们提出了一种基于荧光读数的T7 RNA聚合酶变体的定向进化与2-修饰的核苷酸的接受的筛选测定。通过使用此筛选，我们能够鉴定T2 RNA聚合酶突变体，其2'-甲基硒-2'-脱氧尿苷5'-三磷酸酯的接受度增加。

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《Chemical Communications》 |2012年第79期|p.9870-9872|共3页
作者
Vanessa Siegmund; Tobias Santner; Ronald Micura; Andreas Marx;
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Department of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, Universitaets Strasse 10, 78457 Konstanz, Germany;

Institute of Organic Chemistry, Center for Chemistry and Biomedicine, Leopold-Franzens University, Innrain 80-82, 6020 Innsbruck, Austria;

Institute of Organic Chemistry, Center for Chemistry and Biomedicine, Leopold-Franzens University, Innrain 80-82, 6020 Innsbruck, Austria;

Department of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, Universitaets Strasse 10, 78457 Konstanz, Germany;

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1. Transcription yield of fully 2 '-modified RNA can be increased by the addition of thermostabilizing mutations to T7 RNA polymerase mutants [J] . Meyer Adam J., Garry Daniel J., Hall Bradley, Nucleic Acids Research . 2015,第15期

机译：通过向T7 RNA聚合酶突变体添加热稳定突变，可以提高完全2'-修饰的RNA的转录产量
2. Transcription yield of fully 2′-modified RNA can be increased by the addition of thermostabilizing mutations to T7 RNA polymerase mutants [J] . Adam J. Meyer, Andrew D. Ellington, Bradley Hall, Nucleic acids research . 2015,第15期

机译：通过向T7 RNA聚合酶突变体添加热稳定突变，可以提高完全2'-修饰的RNA的转录产量
3. Efficient synthesis of nucleic acids heavily modified with non-canonical ribose 2′-groups using a mutant T7 RNA polymerase (RNAP) [J] . Robert Padilla, Rui Sousa Nucleic acids research . 1999,第6期

机译：使用突变T7 RNA聚合酶（RNAP）高效合成经非规范核糖2'-基团大量修饰的核酸
4. Inhibitor Candidates's Identification of HCV's RNA polymerase NS5B Using Virtual Screening against iPPI-library [C] . Indah Sulistyawati, Sulistyo Dwi K.P, Mochammad Ichsan International Conference and Workshop on Basic and Applied Sciences . 2016

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5. SINGLE-STRANDED-DNA SP6 AND T7 BACTERIOPHAGE PROMOTER PLASMIDS FOR ENGINEERING MUTANT RNAS AND PROTEINS: EXTENSIONS AND DELETIONS AT THE AMINO TERMINUS WHICH ALTER THE ACTIVITY OF PREPROPARATHYROID HORMONE SIGNAL PEPTIDE. [D] . MEAD, DAVID ALAN. 1986

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6. Transcription yield of fully 2′-modified RNA can be increased by the addition of thermostabilizing mutations to T7 RNA polymerase mutants [O] . Adam J. Meyer, Daniel J. Garry, Bradley Hall, 2015

机译：通过向T7 RNA聚合酶突变体中添加热稳定突变可以提高完全2-修饰的RNA的转录产量
7. Transcribing of Escherichia coli genes with mutant T7 RNA polymerases: stability of lacZ mRNA inversely correlates with polymerase speed. [O] . Makarova, O V, Makarov, E M, Sousa, R, 1995

机译：用突变T7 RNA聚合酶转录大肠杆菌基因：lacZ mRNA的稳定性与聚合酶速度成反比。

Screening mutant libraries of T7 RNA polymerase for candidates with increased acceptance of 2'-modified nueleotidest

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