A random decapeptide library was constructed by using phage-surface display. The oligonucleotide sequence (NNK) was digested with SfiⅠ and NotⅠ and ligated into the phagemid pCANTAB5E. The recombinant DNA was introduced into E.coli TG1 by electroporation, and 5.3×107 phage was harvested. The insert was present in 66.7% of phage,thus the random deca-peptide library had a complexity of 3.53×107.The titer of phage supernatant was 4.8×1011 after the helper phage M13KO7 super-infection. This library was screened using angiogenin protein. 26 ANG-binding clones were indentified from 94 enriched individual phagemid clones after two rounds of panning, The nucleotide sequences encoding peptide recombined in 12 positive phagemid clones were determined. ELISA showed that all of them could specifically bind to ANG.
YANG Hui, ZHANG Ying-Qi, HAN Wei, YAN Zhen, YAO Li-Bo, SU Cheng-Zhi. Construction of Random Decapeptide Library and Screening Identification of Angiogenin-binding Peptides[J]. Progress in Biochemistry and Biophysics,2000,27(6):641-645
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