Selected article for: "expression vector and restriction site"
Author: Funaro, Ada; Gribaudo, Giorgio; Luganini, Anna; Ortolan, Erika; Lo Buono, Nicola; Vicenzi, Elisa; Cassetta, Luca; Landolfo, Santo; Buick, Richard; Falciola, Luca; Murphy, Marianne; Garotta, Gianni; Malavasi, Fabio
Title: Generation of potent neutralizing human monoclonal antibodies against cytomegalovirus infection from immune B cells
  • Document date: 2008_11_12
    • ID: 1u2fkwx3_40
    Snippet: To generate a recombinant IgG1, a consensus 26A1 VH sequence was amplified by PCR with the following primers: 5'-TTTTTTAGATCTCACCATGAACATACTGTGGAG-CATGCTC-3'; 5'-TTTTTTAGATCTTGAGGAGACGGTGA CCAGGGTTCC-3'. The primers contained a BglII restriction site (underlined) for ligation into the hIgG expression vector already containing human IgG1 heavy chain constant domains. The in-house designed bicistronic mammalian retroviral expression vector pRetMEx .....
    Document: To generate a recombinant IgG1, a consensus 26A1 VH sequence was amplified by PCR with the following primers: 5'-TTTTTTAGATCTCACCATGAACATACTGTGGAG-CATGCTC-3'; 5'-TTTTTTAGATCTTGAGGAGACGGTGA CCAGGGTTCC-3'. The primers contained a BglII restriction site (underlined) for ligation into the hIgG expression vector already containing human IgG1 heavy chain constant domains. The in-house designed bicistronic mammalian retroviral expression vector pRetMEx (Fusion Antibodies Ltd. Belfast, Ireland) was a dual promoter vector allowing expression of both antibody chains. The VH domain was cloned into the BamH1 site of multiple cloning site (MCS) 1 of the vector.

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