Selected article for: "dna sequencing and expression vector"
Author: Myllykoski, Matti; Kursula, Petri
Title: Structural aspects of nucleotide ligand binding by a bacterial 2H phosphoesterase
  • Document date: 2017_1_31
    • ID: 0a3okta0_37
    Snippet: Genomic DNA from the BL21(DE3) strain of E. coli was purified and used as a template for PCR (S1 Table) . The initial primers for the first PCR added a TEV cleavage site to the N terminus of the coded protein sequence. A second PCR reaction added attB cloning sites to both ends of the insert. The product from the latter reaction was subcloned into the pDONR221 vector (Invitrogen) and further subcloned into the pTH27 expression vector [34] , which.....
    Document: Genomic DNA from the BL21(DE3) strain of E. coli was purified and used as a template for PCR (S1 Table) . The initial primers for the first PCR added a TEV cleavage site to the N terminus of the coded protein sequence. A second PCR reaction added attB cloning sites to both ends of the insert. The product from the latter reaction was subcloned into the pDONR221 vector (Invitrogen) and further subcloned into the pTH27 expression vector [34] , which adds an N-terminal His 6 tag to the expression product. Clones were verified by DNA sequencing and found to be identical with the LigT database entry (Genbank id. AM946981.2).

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