[Construction,identification and amplification of a yeast two-hybrid random cycle peptide library].

ABSTRACT

AIM: To construct a cycle peptide library composed of 16 random amino acids with yeast two-hybrid system. METHODS: Random oligonucleotides encoding 16-mer peptides were designed and synthesized artificially, and then were amplified by PCR. The amplified products were digested with BamH I and EcoR I and cloned into yeast expression plasmid pGADT(7) GH to construct the cycle library plasmids pGADT(7) GH-RP Then the number of different recombinants and the randomness of the library were tested, and the cycle peptide library plasmids were amplified, extracted and purified. RESULTS: A random cycle peptide library with 1.28 x10(7) different recombinant clones was obtained. No significant difference was found between amino acid distribution in the cycle peptide library and the expected frequency. CONCLUSION: The random cycle peptide library has been successfully constructed. And a lot of cycle peptide library plasmids with high purity were obtained.