Constructs were produced by amplification of complementary oligonucleotides (Integrated DNA Technologies) to produce a full-length sequence containing 5′ Spe I and 3′ Bam HI restriction sites. These were cloned into the vector pJ307 (GST-MBP-His fusion vector) (30 (link)). To distinguish between the termination product and the trans-frame product, we present the MBP tag in the alternative −1 frame relative to the GST tag. MBP and GST were in frame for the positive control. Primers used in this study are provided separately in the Supplementary Materials.
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Synthesis of GST-MBP-His Fusion Constructs
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Corresponding Organization : Stanford University
Other organizations : University College Cork, University of Utah
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Variable analysis
independent variables
- Constructs produced by amplification of complementary oligonucleotides
- Cloning of constructs into the vector pJ307 (GST-MBP-His fusion vector)
- Distinguishing between the termination product and the trans-frame product
- MBP and GST were in frame for the positive control
- MBP and GST were in frame for the positive control
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