Selected article for: "digestion product and MERS cov"
Author: Almazán, Fernando; DeDiego, Marta L.; Sola, Isabel; Zuñiga, Sonia; Nieto-Torres, Jose L.; Marquez-Jurado, Silvia; Andrés, German; Enjuanes, Luis
Title: Engineering a Replication-Competent, Propagation-Defective Middle East Respiratory Syndrome Coronavirus as a Vaccine Candidate
  • Document date: 2013_9_10
    • ID: 14yfs4pa_35
    Snippet: Construction of MERS-CoV cDNA clones lacking accessory genes 3, 4a, 4b, and 5. The deletion of gene 3 was generated by PCR-directed mutagenesis using the plasmid pUC-MERS-1 (a pUC plasmid containing the MERS-1 fragment spanning nucleotides 20,898 to 25,836 of the MERS-CoV genome) as the template and the oligonucleotides MERS-S-Tth111I-VS (5= TGCTATTTGACAAAGTCACTATAGCTGATC 3=, where the restriction site Tth111I is underlined) and MERS-S-PacI-RS (5.....
    Document: Construction of MERS-CoV cDNA clones lacking accessory genes 3, 4a, 4b, and 5. The deletion of gene 3 was generated by PCR-directed mutagenesis using the plasmid pUC-MERS-1 (a pUC plasmid containing the MERS-1 fragment spanning nucleotides 20,898 to 25,836 of the MERS-CoV genome) as the template and the oligonucleotides MERS-S-Tth111I-VS (5= TGCTATTTGACAAAGTCACTATAGCTGATC 3=, where the restriction site Tth111I is underlined) and MERS-S-PacI-RS (5= CCCTTAATTAACTGAGTAACCAACGTCAAAAAGATTCACACT ATTAGTGAACATGAACCTTATGCGGCTCGAGGTCGTATTCC 3=, where the restriction site PacI is underlined). The PCR product, including the deletion (from nucleotides 25,518 to 25,803), was digested with Tth111I and PacI and cloned into the same sites of pUC-MERS-1, leading to pUC-MERS-1-⌬3. To generate pBAC-MERS-⌬3, the SwaI-PacI digestion product from pUC-MERS-1-⌬3 was cloned into the same restriction sites of pBAC-MERS FL (Fig. 3A) .

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