Simultaneous identification of multiple receptors of natural product using an optimized cDNA phage display cloning.
Simultaneously isolating more than one receptor of natural product remains a challenge to chemical genetics. Using cyclosporine A as an affinity probe and an optimized phage display cloning procedure, not only cyclophilin A, but also cyclophilin B was isolated as the full-length gene clone from a human brain cDNA library. This optimized protocol can be used to select protein targets of chemicals dependent on the binding affinity rather than on the relative abundance in cells.
State Key Laboratory of Bio-organic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 354 Fenglin Road, Shanghai 200032, China.
Qing-Li
QL
He
Hui
H
Jiang
Feng
F
Zhang
Hai-Bao
HB
Chen
Gong-Li
GL
Tang
D001435:Bacteriophages / Q000378:metabolism; D019921:Biotinylation; D001921:Brain / Q000378:metabolism*; D002626:Chemistry, Pharmaceutical / Q000379:methods; D003001:Cloning, Molecular; D021984:Cyclophilin A / Q000737:chemistry; D021983:Cyclophilins / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D015723:Gene Library; D006801:Humans; D007700:Kinetics; D008956:Models, Chemical; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D019151:Peptide Library; D013678:Technology, Pharmaceutical / Q000379:methods
D018076:DNA, Complementary; D019151:Peptide Library; C081786:cyclophilin B; D021984:Cyclophilin A; D021983:Cyclophilins