201 X2CX14CX2+X2CX18 phage display library pool 18, 20 Erkki Koivunen (Division of Biochemistry, University of Helsinki, Helsinki, Finland) Semi-random NNK 202 X3YX4 phage display library 7 3e8 Hermann Gram Semi-random NNK Linear The X3YX4 phage display library is based on the phagemid vector pGEM-gIII, displaying randomized peptides fused to gpIII on the surface of M13 filamentous phage. The peptide library was randomized at seven positions surrounding an invariant tyrosine residue. 203 X5YSKPPPIP M13 phage display library 5 Leslie J. Berg Completely random Linear 204 X2CX4-8CX2 phage display library pool 10-14 3e8 De Ciechi Semi-random NNK Circular Five constrained libraries wherein random peptides, varying from four to eight amino acids, are flanked by a Cys on either side in addition to a further two random residues on either side were constructed with pMON20401 vector, which was derived from fUSE3 vector. The peptides are expressed immediately following the gene III signal peptide cleavage site. The codons of the degenerate oligonucleotides used for construction of the libraries followed the formula NNK, where N is an equimolar mixture of all four deoxynucleotides and K is an equimolar mixture of dGTP and dTTP. NH2-X2CX4-8CX2-GAAGGAGAGAG=TVES... 205 X10+X9GAX9 phage display library pool 10,18 2e8, 4e8 De Ciechi Semi-random Linear The 10-mer and 20-mer linear library were constructed with vector pMON6000 and pMON20401 respectively. Both vectors were derived from the fUSE3 vector. The peptides are expressed immediately following the gene III signal peptide cleavage site. The codons of the degenerate oligonucleotides used for construction of the libraries followed the formula NNK, where N is an equimolar mixture of all four deoxynucleotides and K is an equimolar mixture of dGTP and dTTP. NH2-X10-GG=TVES; NH2-X9GAX9-GAAGGAGAGAG=TVES...... 206 CX6C phage display library 6 C. Demangel Completely random Circular This library is constructed with the expression vector pC3H, which is derived from the phagemid pBluescript designed by Stratagen. A collection of oligonucleotides encoding random hexapeptides flanked by two cysteines and four constant residues EDGACXXXXXXCGAAS was inserted in the XhoI-SpeI cloning site. The insert was located between a leader sequence (pelB) and a truncated gene III, and allowed the monovalent display of random peptides in a constrained form on the N-terminus of the protein III. 207 X8 phage display library 8 Angus C. Nairn Completely random NNS Linear This library was constructed with vector M13CL2, which was derived from the phage vector M13mp18. Random peptide sequences were expressed on the N-terminus of the minor coat protein (gpIII). NH2-AX8GAAGAIEGR=AETV...GAAGA is the flexible-arm sequence and IEGR is the recognition sequence for Factor Xa. 208 X20 phage display library 20 1e8 Jonathan M. Gershoni Completely random NNK Linear This library was constructed in the fUSE5 vector. A 93 nucleotide long oligonucleotide encoding random 20 amino acid sequences was inserted into gene III at the region corresponding to the amino terminus of pIII. The oligonucleotides ON93 (5'-GAGCCAGTGCATCA(NNK)20TCGCTAACAGGTGGGTCTG-3') containing the degenerate sequences NNK20. 209 X10 fUSE5 phage display library 10 4.0e8 Philippe Valadon Completely random NNK Linear This decapeptide library was based on the fUSE5 vector. Random peptides were displayed at the N-terminal end of the pIII coat protein of the tetracycline-resistant phage fd. 210 CX7C and CX9C phage display library pool 7, 9 Erkki Koivunen (Division of Biochemistry, University of Helsinki, Helsinki, Finland) Completely random NNK Circular The phage librarie were based on the fUSE5 vector. 211 X6 phage display library 6 Klaus Mosbach Completely random Linear pIII protein NH2-ADGAX6GTAG 212 D38 phage display library 35 5e8 Stephen J. McConnell Semi-random NNB This long peptide library was constructed in an M13 cloning vector (CYT-V1) that was generated by modifying the RF form of vector M663. The original M663 was derived from M13mp. The N-terminal pIII fusion proteins displayed in the D38 library are X20[YHND]A[IMTNKSR]X15. X residue is encoded by NNB. 213 DC43 phage display library 40 1e8 Stephen J. McConnell Semi-random This long peptide library was constructed in an M13 cloning vector (CYT-V1) that was generated by modifying the RF form of vector M663. The original M663 was derived from M13mp. The N-terminal pIII fusion proteins displayed in the DC43 library are X20GCGX20. X residue is encoded by NNB. 214 X10 phage display library 10 2e8 De Ciechi Completely random NNK Linear The 10-mer linear library was constructed with vector pMON6000 and presented on fd-tet. The peptides are expressed immediately following the gene III signal peptide cleavage site. NH2-X10-GG=TVES... 215 R8C phage display library 8 1.1e8 Heather Hanson Pierce Completely random Linear Two oligonucleotides were synthesized to generate the DNA cassette used in the construction of the conformationally constrained library. The first was a degenerate oligonucleotide, 5'-TGACGTCTCGAGTTGT(NNK)8-TGTGGATCTAGAAGGATC-3', containing an Xho I site, where N represents an equimolar mixture of A, C, G and T; and K corresponds to an equimolar mixture of G and T. The NNK coding scheme utilizes 32 codons to encode 20 amino acids; the frequency of each amino acid is once (Cys, Asp, Glu, Phe, His, Ile, Lys, Met, Asn, Gln, Trp, Tyr), twice (Ala, Gly, Pro, Val, Thr), or thrice (Leu, Arg, Ser) per codon. The second oligonucleotide, 3'-CCTAGATCTTCCTAG-5', contained an Xba I site, and served as a primer for DNA synthesis. 216 BPTI/Delta-g3p/biased 8-mer phage display library 4 6.2e7 Steven P. Weinheimer Completely random NNS Linear This substrate library was based on the phagemid vector pCANTAB5E. GGPGG-YLQAX4-GGPGG. The sequence YLQA is from the HSV-1 wild-type R cleavage site. YLQAX4 was flanked on either side by the amino acids GGPGG to disrupt any neighboring secondary structure that might influence protease cleavage. 217 BPTI/Dg3p/6-mer phage display library 6 6e8 Steven P. Weinheimer Completely random NNS Linear This substrate library was based on the phagemid vector pCANTAB5E. GGPGG-X6-GGPGG. The randomized insert was flanked on either side by the amino acids GGPGG to disrupt any neighboring secondary structure that might influence protease cleavage. 218 X10 phage display library 10 4e8 Christian (chiron) Completely random NNK Linear The random peptides were expressed on the protein PIII of bacteriophage fd (PMID: 1404385). 219 X8 phage display library 8 2e7 Joerg Koehl Completely random NNK Linear The library was based on the vector pCANTAB5 (Pharmacia). Random peptides were expressed on the protein PIII. 220 CWl phage display library 12 1e9 Naohiko Ikegaki Completely random NNK Linear The CW1 library is an X12 library with NNK coding scheme. The random peptides are displayed at the N-terminus of mature PIII. The library was constructed in a bacteriophage Ml3 vector, mBAX. 221 X21 phage display library 21 6.5e7 Anthony J. Conley Completely random NNK Linear The library was based on the vector fUSE5. Random peptides were expressed on the protein PIII. 222 CX5GPXRX5C bias phage display library 11 Anthony J. Conley Semi-random NNK Circular The library was based on the vector fUSE5. GPXR represents the V3 region apex sequence. Random peptides were expressed on the protein PIII. 223 LLX5GPXRX5LL bias phage display library 11 Anthony J. Conley Semi-random NNK Linear The library was based on the vector fUSE5. GPXR represents the V3 region apex sequence. Random peptides were expressed on the protein PIII. 224 CX5C phage libray 5 Erkki Koivunen (Division of Biochemistry, University of Helsinki, Helsinki, Finland) Completely random Circular 225 X16 phage display library 16 Valery Alakhov Completely random NNK Linear The library containing 16-amino acid peptide was constructed , using fUSE5 as the phage vector.