BDB - Library Details

Library

Library Name: pGWX4YX4 phage display library

Library ID:

298

Library Length:

Library Complexity:

1.75e8

Library Titer:

Library From:

Su-Jun Deng, Gaochao Tian

Library Randomness:

Semi-random

Library Scheme:

NNK

Library Topology:

Linear

Comments:

The library was constructed in the vector pGWg3. To construct phage display vector pGWg3, EcoRI- and HindIII-digested pUC18 was ligated with the M13 phage gIII fragment after being amplified using PCR with oligonucleotides 5’-GGGGAATTCA AACTCAGATC TGGTGGCGGT GGATCCCCAT CGTTTGTGAA TATCAAGGC-3’ and 5’-GCCAAGCTTC TAATAATAAC GGAATACCCA AAAGAACTG-3’. Subsequently, the small PvuII fragment of the ligated product that contained gIII was inserted into the large fragment of PvuII-digested pBluescript. A DNA fragment encoding Pel B leader peptide and multiple cloning sites was constructed by a synthetic ligase chain reaction approach using 4 oligonucleotides 5’-AATTGAATAG AGGGTAGAAT TGAAATACCT GCCGACCGCT GCTGCTGGTC TGCTCCTCGC GGCCC-3’, 5’-AGCCGGCCAT GGCCCAACTC GAGTGGGCGG CCGCAGGTGG ATAACAGAAA CTGATCTCCG AAGGGTACCA AGACCTGAAC A-3’, 5’-GATCTGTTCA GGTCTTGGTA CCCTTCGGAG ATCGTTTCT GTTATCCACC TGCGGCCGCC CACTCGAGTT GGGCCA-3’, and 5’-TGCCGGCTGG GCCGCGAGGA GCAGACCAGC AGCAGCGGTC GGCAGCAGGT ATTTCATCAA TTCTACCCTC TATTC- 3’. Insertion of this DNA fragment into the EcoRI- and BglII-digested pBluescript containing gIII yielded the phage display vector, pGWg3.

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