pGL3-Basic Vector

Protocol tips

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	Cut vector between KpnI-BglII fragments Insert 0.3 kb and 0.17 kb DNA fragments

Protocol tips
Cut vector between KpnI-BglII fragments Insert 0.3 kb and 0.17 kb DNA fragments

Publication protocol

In our previous study, the reporter plasmids, pGL3-0.3 and pGL3-0.17, in which the promoter regions −253/+47 and −122/+47 of the β4GalT4 gene relative to the transcriptional start site were inserted into the firefly luciferase reporter vector, pGL3-Basic (Promega, Madison, WI, USA), were constructed [10]. To establish the stable sensor cells having the luciferase gene under the control of the β4GalT4 gene promoters from SW480 cells, two reporter plasmids containing 0.3 kb and 0.17 kb promoter regions were prepared using pGL4.15[luc2p/Hygro] vector (Promega), which contains hygromycin-resistant gene. In brief, after the KpnI-BglII fragments were excised from pGL3-0.3 and pGL3-0.17, the 0.3 kb and 0.17 kb DNA fragments were inserted between KpnI and BglII sites of pGL4.15[luc2p/Hygro] vector to generate pGL4-0.3 and pGL4-0.17, respectively.

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Manufacturer protocol

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