The emergence of drug-resistant protease(PR) has seriously affected the anti AIDS therapy. Using mutated PR to screen phage library displaying randomized HIV PR target sites, phages susceptible to mutated PR can be obtained ,and used in drug screening of protease inhibitor(PI) against to drug-resistant HIV PR. In order to validate the feasibility of this suppose, a designed LD3-CAP2NC peptide composing of HIV CAP2NC with P2/NC target site and LD3 peptide locating in the NH2 terminus of the CAP2NC which could immobilise the phage to the plate through binding to human IgG was displayed on the surface of phage. This phage was fixed on plate, and suffered to the cleaving by HIV SF2 PR. The uncleaved phage leaving on the plate was measured using HRP/Anti-M13 conjugate, which reflects the cleavage efficiency. The results showed the phage can be cleaved effectively in a dose-dependent manner to PR concentration, with the most cleavage effect reached to more than 80%.Comparing with the control, the ELISA value of the cleaved phage decreased obviously. Furthermore, this cleavage was specifically inhibited by HIV protease inhibitor durg Indinavire. The data proved that a novel HIV protease cleavage model of the phage displaying Gag CAP2NC was firstly successfully established, which can not only be used as a new research platform to investigate the relationship between PR drug-resistant mutation and PR target sequence adapted mutation, but also lay a foundation to establish the new phage screening model for PIs especially against to drug-resistant PR.