酵母双杂交随机环肽库的构建、鉴定与扩增  

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

作　　者：徐祥[1] 梁华平[1] 王付龙 罗艳[1] 王正国[1] 

机构地区：[1]第三军医大学附属大坪医院野战外科研究所一室,重庆400042

出　　处：《细胞与分子免疫学杂志》2003年第5期437-439,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家重点基础研究规划 (973)资助(No .G1 9990 542 0 3);国家自然科学基金资助项目(No .30 0 80 0 0 9);重庆市科委基金资助项目(No.2 0 0 0 631 9)

摘　　要：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。AIM: To construct a cycle peptide library composed of 16 random amino acids with yeast two hybrid system. METHODS: Random oligonucleotides encoding 16 peptides were designed and synthesized artificially, and then random oligonucleotides were amplified by PCR.The amplified products digested with Bam H I and Eco R I were 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 identified , and the cycle peptide library plasmids were amplified, extracted and purified.RESULTS: A random cycle peptide library with 1.28×10 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.

关 键 词：随机环肽库 酵母双杂交 pGADT7GH 构建 鉴定 扩增 

分 类 号：R373.9[医药卫生—病原生物学] R341.43[医药卫生—基础医学]