Methods of screening molecular libraries for active molecules that restore the function of target proteins and peptides that restore the function of mutant p53

摘要

The present invention provides a method of screening a library of peptides of formula M-G/M/V-(X)n wherein n is an integer from 3 to 18, M is methionine, G is glycine, V is valine and each X, which may be the same or different, is any genetically encoded amino acid, which method comprises: a) transforming a host cell population with a library of nucleic acid constructs which can express free peptides of formula M-G/M/V-(X)n; b) culturing the transformed host cells under conditions suitable for intra-cellular expression of the peptides of formula M-G/M/V- (X)n; and C) analysing the host cell population to determine the effect of the peptides of formula M-G/M/V- (X) n on a reporter system. Also describe are a library of nucleic acid constructs which can express free peptides in an intra-cellular environment, said peptides having the sequence M-G/M/V-(X)n, wherein n is an integer from 3 to 18, M is methionine, G is glycine, V is valine and each X, which may be the same or different, is any genetically encoded amino acid; and a method of generating a library of nucleic acid constructs which can express free peptides in an intra-cellular environment, which method comprises synthesising a library of DNA molecules which include the nucleotide sequence ATGGGA(NNK)n, wherein n is an integer from 3 to 18, N is A, C, T or G and K is G or T and wherein each NNK triplet may be the same or different, and inserting a library of synthesised DNA fragments which each includes a nucleotide sequence of formula ATGGGA(NNK)n into expression vectors.