Method and its use to measure the stability of the protein

摘要

The present invention provides a genetic construct which structure is represented by 5'-A + B + C + D + E-3 '. Here, A is a promoter, B is a coding sequence for a fusion protein consisting of the protein of interest and a first labeled protein, the coding sequence of the C is the connecting peptide, D is the coding sequence of the second labeled protein , and E represents the terminator. Further, the present invention provides a polypeptide structure is represented by B1 + C1 + D1. Here, B1 is a fusion protein consisting of the protein of interest and a first labeled protein, C1 is the connecting peptide, and the D1 represents the coding sequence of the second labeled protein. Furthermore, the present invention provides a vector comprising said genetic construct, to provide the genetic construct or a mammalian cell comprising a vector, and methods and applications for measuring the stability of the constructed library as well as protein in said cells. The method of the present invention not only has high sensitivity and specificity for the measurement of protein stability, the operation is simple.