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.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
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)~~
