BDB - Library Details

Library

Library Name: Aptide pIGT2-based phage display library (X6GSWTWENGKWTWKGX6)

Library ID:

471

Library Length:

Library Complexity:

8e8

Library Titer:

Library From:

Sangyong Jon (Gwangju Institute of Science and Technology, Gwangju, South Korea)

Library Randomness:

Semi-random

Library Scheme:

NNK

Library Topology:

Circular

Comments:

An aptide comprises a stabilizing scaffold and two target-binding regions. The scaffold consists of a small (12 amino acids) but highly stable tryptophan zipper that forms a leucine-zipper-like b-hairpin structure, in which two tryptophan–tryptophan cross-strand pairs create a robust and stable structure. To mimic the DNA recognition site of bZIP proteins two target-binding regions, each comprising six randomizable amino acids, are introduced at both ends of the trpzip scaffold through glycine linkers. To create a large and highly diverse aptide library, we synthesized two degenerate aptide-encoding oligonucleotides: aptide-F1, 5'-TTC TAT GCG GCC CAG CTG GCC (NNK)6 GGA TCT TGG ACA TGG GAA AAC GGA AAA-3 and aptide-B1, 5’-AAC AGT TTC TGC GGC CGC TCC TCC TCC (MNN)6 TCC CTT CCA TGT CCA TTT TCC GTT-3’, where N was A, T, G, or C; K was G or T; and M was C or A (Genotech). A double-stranded randomized insert was created using polymerase chain reaction (PCR), double-digested with SfiI/NotI (New England Biolabs) and cloned into SfiI/NotI-digested pIGT2 phagemid vectors (Ig Therapy Co.). The resulting construct was electroporated into electrocompetent E. coli XL1 cells (Stratagene), and the transformed cells were infected with Ex12 mutant helper phage (Ig Therapy Co.) for amplification of the aptide-displaying phage library.

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