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


CONCLUSIONS
The random cycle peptide library has been successfully constructed. And a lot of cycle peptide library plasmids with high purity were obtained.
Xu
Xiang
X
Research Institute of Surgery, Third Military Medical University, Chongqing 400042, China. xuxiang75@cta.cq.cn
Liang
Hua-ping
HP
Wang
Fu-long
FL
Luo
Yan
Y
Wang
Zheng-guo
ZG
D000595:Amino Acid Sequence; D001483:Base Sequence; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics*
D019151:Peptide Library