Selected article for: "Applied Biosystems flight mass time spectrometer and protein concentration"
Author: Amrita Roy; Liangzhong Lim; Shagun Srivastava; Jianxing Song
Title: Unique properties of Zika NS2B-NS3pro complexes as decoded by experiments and MD simulations
  • Document date: 2016_9_28
    • ID: l82aakrk_52
    Snippet: Plasmid construction-The identified genes encoding NS3pro (1-185) and NS2B (1-130) from the Asian Zika strain 8375 (GenBank ID: KU501217.1) were optimized and synthesized by GenScript (Piscataway, NJ). With designed primers, the genes by GenScript were used as templates for amplifying DNA fragments encoding the isolated NS3 (14-185) and NS2B (48-100) with the transmembrane regions removed, as well as NS2B (48-74) with the C-region further deleted.....
    Document: Plasmid construction-The identified genes encoding NS3pro (1-185) and NS2B (1-130) from the Asian Zika strain 8375 (GenBank ID: KU501217.1) were optimized and synthesized by GenScript (Piscataway, NJ). With designed primers, the genes by GenScript were used as templates for amplifying DNA fragments encoding the isolated NS3 (14-185) and NS2B (48-100) with the transmembrane regions removed, as well as NS2B (48-74) with the C-region further deleted. Furthermore, DNA fragments were designed for linking NS2B (48-100) to NS3 (14-185) by (Gly)4-Ser-(Gly)4 using overlap PCR as previously described (14) . Amplified DNA fragments were subsequently cloned into His-tagged pET28a vector (Novagen). DNA sequences of all constructs were verified by automated DNA sequencing. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/078113 doi: bioRxiv preprint and protein sequencing with time-of-flight-mass spectrometer (Applied Biosystems). Protein concentration was determined by the UV spectroscopic method with 8 M urea (12) .

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