酵母双杂交随机肽库的设计及构建  

Design and construction of a random peptide library with yeast two hybrid system

作　　者：胡承香[1] 徐祥[1] 梁华平[1] 吕凤林[1] 王付龙 王正国[3] 

机构地区：[1]第三军医大学大坪医院野战外科研究所第一研究室 [2]河北省邯郸市解放军第285医院外一科,邯郸056001 [3]第三军医大学大坪医院野战外科研究所第四研究室,创伤烧伤复合伤国家重点实验室,重庆400042

出　　处：《第三军医大学学报》2004年第1期61-63,共3页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划资助项目 ("973"项目 ) (G19990 54 2 0 3) ;国家自然科学基金资助项目 ( 30 0 80 0 0 9) ;重庆市科委基金资助项目( 2 0 0 0 6 319)~~

摘　　要：目的　构建一个含 16个氨基酸的酵母双杂交随机肽库。方法　设计合成编码 16肽的随机DNA片段 ,PCR扩增随机DNA片段 ,经限制性内切酶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肽库质粒pGADT7GH RP并检验其库容。结果　扩增获得编码 16肽的随机DNA片段 ,并成功将随机DNA片段克隆入酵母表达质粒pGADT7GH ,与GAL4AD形成融合蛋白 ,其库容为 1 2 8× 10 7。Objective To construct a peptide library composed of 16 random amino acids with yeast two hybrid system. Methods Random oligonucleotides encoded with 16 peptides were designed and artificially synthesized, and were amplified by PCR. The amplified products after being digested with Bam HⅠ and Eco RⅠ were cloned into plasmid pGADT7GH to construct the library plasmids pGADT7GH RP. Results The random oligonucleotides encoded with 16 peptides were obtained and inserted into pGADT7GH, and became fusion protein with GAL4AD. Conclusion The random peptide library with 1 28×10 7 different recombinant clones has been successfully constructed.

关 键 词：随机肽库 酵母双杂交 pGADT7 GH 

分 类 号：Q516[生物学—生物化学] Q503