BDB - Library Details

Library

Library Name: X6 T7 phage display library

Library ID:

463

Library Length:

Library Complexity:

2.5e6

Library Titer:

Library From:

Kazuhisa Sugimura (Kagoshima University, Korimoto, Japan)

Library Randomness:

Completely random

Library Scheme:

NNM

Library Topology:

Linear

Comments:

The T7 phage library displaying random 6-mer peptides was constructed using the phage vector T7Select415 (Novagen). To add hexameric amino acid sequences to the C-terminal of the G10 protein of T7 phage through the GGGS linker peptide, a template oligonucleotide (5'-GCCGCAAGCTTTTATCCMNNMNNMNNMNNMNNMNNCGAACCTCCACCTGAATTCGG-3') was synthesized. This oligonucleotide was amplified by PCR using forward and reverse primers (5'-CCGAATTCAGGTGGAGG-3'and 5'-CGGCGTTCGAAAATAGG-3') that harbored restriction sites for EcoRI and HindIII, respectively. The generated PCR product was purified on a PCR-M column (Viogene) and digested with EcoRI and HindIII. The DNA fragment was purified on a PCR-M column again and ligated into the corresponding restriction sites in the T7Select415 vector. The ligation mixture was subjected to anin vitro packaging reaction using a T7 packaging extract (Novagen). The packaged T7 phages were infected into Escherichia coli BL21 and amplified once.

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