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. 1999 Oct 12;96(21):11723–11728. doi: 10.1073/pnas.96.21.11723

Figure 6.

Figure 6

Model selection of scFvF8-VP16 clone from a mixture of 500,000 scFv clones. Yeast L40 was cotransformed with the AMCVp41/BTM116 DNA-binding bait clone together with DNA of a nonrelevant, naive scFv-VP16 library into which scFv-F8-VP16 had been added at a dilution of 1 in 500,000. Yeast were plated on plates lacking histidine to select for interacting partners. Forty eight clones that grew on these plates were picked and grown on a fresh grid and the β-gal assay was performed (a). Ten blue clones were grown, DNA was prepared, and the fingerprint of BstN1 digestion products compared with scFv-F8-VP16. All 10 clones showed an indical pattern to this clone (b). DNA sequencing (not shown) confirmed that the selected clones were identical to scFv-F8-VP16. (a) Forty eight colonies from his− plates were restreaked and β-gal was assay carried out. Twenty two colonies (46%) showed a robust blue coloration. (b) Ten of these colonies were picked into liquid culture and plasmid DNA was prepared after 2 days growth. The plasmid DNA were digested with BstN1 and separated on agarose in comparison with scFv-F8-VP16. Sizes are indicated from coelectrophoresis of size markers and indicated on the left.