BDB - Library Details

Library

Library Name: TN10 phage display library

Library ID:

355

Library Length:

Library Complexity:

Library Titer:

Library From:

Dyax Corp

Library Randomness:

Semi-random

Library Scheme:

Library Topology:

Circular

Comments:

The disulfide-constrained cyclic peptide phage display library TN6 was constructed in a derivative of M13mp18, MANP, having the following modifications: bla (AmpR) gene, modified junction between signal of iii and coding region for mature III, and removal of LacZ complementation system. MANP comprises 8164 bases. The bla gene was from pGEM3Zf1(+), was bound by BamHI-HindIII sites at the 5' end and HindIII-SalI sites at the 3' end, replaced bases 6001 through 6432 of M13mp18, and was in the same orientation as the phage genes. The junction between the III signal sequence and mature III was modified with an NcoI restriction site embedded in the last three codons of the signal sequence. After the III signal, MANP contained the sequence of BPTI with restriction sites for the enzymes StyI, XhoI, PflMI, ApaI, Bsp120I, EcoO109I, PspOMI, BssHII, StuI, BstZ17I, BlpI, EagI, and SphI, which were all unique within MANP. Following BPTI, a unique PstI site and a Factor Xa cleavage site were inserted in a short linker region before mature III. The DNA encoding the library was synthesized with constant DNA on either side so that the DNA could be PCR amplified using TAQ DNA polymerase, cleaved with NcoI and PstI, and ligated to similarly cleaved vector. The variegated parts were synthesized with TRIM technology, which incorporates trinucleotides and allows mixtures of any set of amino acid types in any desired proportions. The varied DNA sequence positions of TN10 phage library is depicted as "X8X8X2 CX1X1X1X1X1X1X1X1C X2X8X8", where X1=ADFGHLNPQRSVWY, X2=ADEFGHIKLMNPQRSTVWY and X8=DFHLNPRSWY.

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