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目的 :构建含 16个氨基酸的酵母双杂交随机环肽库。方法 :以PCR扩增人工合成的随机DNA模板 ,经BamHI和EcoRI酶切后 ,克隆入酵母表达质粒pGADT7GH ,构建酵母双杂交随机环肽库并检验其库容及随机性 ,扩增、抽提并纯化酵母双杂交环肽库质粒。结果 :构建了酵母双杂交随机环肽库 ,库容为 1.2 8× 10 7,氨基酸分布与理论频数无显著差异 ,并扩增获得大量高纯度的环肽库质粒。结论 :成功地构建酵母双杂交随机环肽库 ,并获得大量高纯度的环肽库质粒。
Objective: To construct yeast two-hybrid random peptide library containing 16 amino acids. Methods: Random amplified DNA template was amplified by PCR, digested with BamHI and EcoRI, and cloned into yeast expression plasmid pGADT7GH to construct a yeast two-hybrid random peptide library. The random amplified polymorphic DNA Purification of yeast two-hybrid cyclic peptide library plasmid. Results: The yeast two-hybrid random cyclic peptide library was constructed with a storage capacity of 1.2 8 × 10 7. There was no significant difference in amino acid distribution and theoretical frequency, and a large number of high purity cyclic peptide library plasmids were obtained. Conclusion: The yeast two-hybrid random peptide library was successfully constructed and a large number of plasmids with high purity were obtained.