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Figure 1 | BMC Biotechnology

Figure 1

From: Creation and validation of a ligation-independent cloning (LIC) retroviral vector for stable gene transduction in mammalian cells

Figure 1

Schematic depicting the conversion of the pBABE-Neo plasmid into pBLIC-Neo by insertion of a specially designed LIC adaptor. The LIC adaptor consists of a PmlI site and a 12/13 base pair sequence used for LIC cloning (for generating overhangs consisting of 13 bases on the left side and 12 bases on the right). The PmlI restriction sequence is bolded and the digestion site is indicated by an inverted triangle within the restriction sequence. The terminal overhang sequences for BamHI (right) and SalI (left) that allow ligation of the LIC adaptor into the original pBABE-Neo plasmid are italicized. Note that the palindromic nature of the SalI site (G/TCGAC) will be lost upon ligation of the adaptor sequence with the digested pBabe plasmid (CTCGAC). The guanines that serve as T4 exonuclease activity termination sites are bolded and italicized.

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