76 AA-xxRx(K/R)Rx-AA phage display library 7 James A. Wells Semi-random Linear 77 AA-xxRxPRx-AA phage display library 7 James A. Wells Semi-random Linear 78 X10 phage display library 10 Matthew D. Scharff Completely random NNK Linear The library is based on fUSE5 79 L200 phage display library 19 Matthew D. Scharff Semi-random Linear The L200 library was created by inserting a modified version of the PA1 motif between the SfiI sites of the vector fUSE5, located at the amino-terminal part of the pIII coat protein giving the final amino acid sequence H2N-ADVA X6 TPXW[M/L][M/L] X6 AAG-g3p. 80 CX(7-10)C and X(9-10) phage display library pool 7-10 Erkki Koivunen (Division of Biochemistry, University of Helsinki, Helsinki, Finland) Completely random 81 Three CX12C phage display library pool 12 Jonathan M. Gershoni Completely random Circular Each library represented 1e9 - 5e9 recombinant random 12mer peptides flanked by constant cysteine residues so as to constrain a looped structure at the NH2 terminus of the major coat protein, pVIII, of the folamentous bacteriophage fd. Three 12mer cysteine-constrained peptide libraries were constructed: one in ftac88 and the other two in fth1. 82 CX12C phage display library 12 Jonathan M. Gershoni Completely random Circular 83 X6PX6 and X6YX6 phage display library pool 12 Mark W. Grinstaff Semi-random NNK Linear 84 CX6C phage display library 6 6.4e7 1.1e9 Indraneel Ghosh Completely random NNS Circular The six-residue disulfide-constrained cyclic peptide library was constructed N-terminal to a peptide linker to the gene III fusion protein encoded by the phagemid vector pCANTAB-5E (Amersham Biosciences, Princeton, NJ, USA). A gene encoding a peptide linker and containing an internal PstI restriction site had been previously cloned into pCANTAB-5E between SfiI and NotI restriction sites in our laboratory to produce pCANTAB-Fos. 85 CX7C T7 phage display library 7 1.28e9 3.3e7 University of Tartu, Tartu, Estonia Completely random NNK Circular This library was constructed using the T7 select-415 kit from Novagen with NNK codon. Target peptides were expressed as a fusion to the C-terminus of the 10B capsid protein and were displayed on the virion surface where they were accessible for interaction with other proteins or ligands. The displayed peptide was situated between cysteine residues and therefore, formation of a disulphide bridge would join the ends of the heptapeptide. The fusion polypeptide was present in 415 copies on each phage particle. 86 X12 phage display library 12 Jonathan M. Gershoni Completely random Linear 87 Ph.D.-12, Ph.D.-7 and Ph.D.-C7C phage display library pool 7, 12 New England BioLabs, Beverly, MA, USA Completely random NNK 88 X5-Fixed-X5 phage display library 10 James R. Burke Completely random Linear An assortment of libraries representing different fixed residue and other structural constraints will provide the greatest likelihood of deriving high-affinity peptides specific for a given target. 89 fUSE5-based CX7C phage display library 7 2e8 1e12 Renata Pasqualini (University of Texas, M. D. Anderson Cancer Center, Houston, TX) Completely random Circular 90 Type 8 phage display library (X8) 8 Angela M. Belcher Completely random NNM Linear The M13KE phage vector was modified by making a cloning site for pVIII display. A Pst I restriction site was made by mutating T to A at position 1372, a BamH I site was made by mutating C to G at positon 1381, and the Pst I site at position 6246 was deleted by mutating T to A at position 6250. The site-directed mutagenesis was done using overlap extension PCR. A dsDNA library was then prepared and cloned into the resulting modified phage vector, named M13SK, using Pst I and BamH I. To obtain the dsDNA library, partial library duplexes were formed by annealing of extension primer (5\'-GATGCTGTCTTTCGCTGCAG-3\') with oligonucleotides (3\'-ACGACAGAAAGCGACGTCnm(nnm)6nnCCTAGGAACATC ATC-5\', where n=A, T, C, or G and m=A or C). The partial library duplexes were incubated with Klenow fragment (3\'→'5\' exo-) (10 U/μL) and dNTP at 37 °C for 30 min. The Klenow fragment was inactivated by heating (75 °C for 20 min), and the mixture was digested with Pst I and BamH I. The digested DNA was gel purified (2-40% TBE polyacrylamide gel), ligated into M13SK, and transfected to XL1-Blue Electroporation Competent Cells using a MicroPulser (Biorad). The library was titered according to manufacturer directions and sequenced (MIT Biopolymers Laboratory) before amplification. 91 pSKAN8-HyA phage display library 8 MoBiTec GmbH (Gottingen, Germany) Completely random Circular The variable region is a 8-aa (pSKAN8-HyA) extended peptide held between two disulfide bridges at the exposed tip of the human pancreatic secretory trypsin inhibitor (PSTI). A phagemid pSKAN8 was created which contains a fusion between the PSTI and M13 pIII protein-coding genes. 92 X12 phage display library 12 Antonio Verdoliva Completely random Linear The library was based on pC89. 93 X12 phage display library 12 Nicholas M. Boulis Completely random Linear 94 CX7C phage display library 7 MoBiTec GmbH (Gottingen, Germany) Completely random Circular 95 X4YX4 phage display library 8 T.S. Pillay Semi-random Linear The library was constructed by subcloning oligonucleotides encoding a random nonapeptide sequence into the 10B gene of the icosahedral T7 bacteriophage, using the T7 Select 415-1b cloning kit (Novagen). The cloned oligonucleotide, 5'-(NNB)4-TAT-(NNB)4-3', of the synthetic construct contained a conserved tyrosine (codon TAT) residue critical for phosphorylation at the center. 96 X6PPX(Y/F)X6 phage display library 13 Bernard Weisblum Semi-random Linear phage display library was based on insertion of a random DNA sequence, as indicated, in a multiple cloning site at the N-terminus of phage M13 protein pIII 97 CX9C and CX15C phage display library pool 9, 15 M. A. Persson Completely random Circular 98 X12 phage display library 12 Yuan Wang and You-hua Xie Completely random Linear The gene for the pVIII protein of phage M13 was amplified by PCR with VCSM13 (Stratagene, La Jolla, CA) as the template and inserted into pCANTAB5E (Amersham Pharmacia, Uppsala, Sweden), replacing the original gene III. The new vector, designated pFuse8, was used as parental vector for pVIII-based phage display. 99 CDX3KPCALLRYX10 phage display library 13 Igor Fisch Completely random Circular 100 LX-8, X15, X8CX8, X30 phage display library pool 12, 15, 17, 30 J. K. Scott (Simon Fraser University, Burnaby, Canada) Semi-random This is a mixture phage display library (LibraryID: 43-45,50).