Method of target-integrating foreign DNA (Deoxyribonucleic Acid) sequence to Rosa26 sites of rat and mouse as well as application thereof
A DNA sequence, exogenous technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., to achieve the effect of clear genetic background, accurate biological function, and shortened screening period
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[0040] 1. Rat and mouse TALENs recognize the Rosa26 site
[0041] After research, it was found that a TALEN that can be used for targeted integration of foreign DNA sequences into rats and mice recognizes the Rosa26 site. The upstream and downstream recognition sequences of the TALEN that recognizes the Rosa26 site are as follows:
[0042]Upstream recognition sequence: 5'-agccaataatcaaatt-3' (as shown in SEQ ID NO.1)
[0043] Downstream recognition sequence: 5'-agttccttggtaacatt-3' (as shown in SEQ ID NO.2).
[0044] where the details are available as figure 1 shown.
[0045] 2. Construction of a pair of TALEN plasmids that recognize the above sites
[0046] 1. TALEN recognizes the upstream and downstream TALEN sequence synthesis and digestion of the Rosa26 site
[0047] The following upstream and downstream TALEN sequences (CDS coding regions) were chemically synthesized with BsmB I sites at both ends. For the synthesized sequence, the enzyme BsmB I was used to digest, a...
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