OBJECTIVE To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus. METHOD Using primer Premier 5.0 software, two pairs of nested PCR primers were designed to amplify the N gene. After purification, the amplified products were cloned into pMD18-T vectors, and the positive clones with the inserted fragments were identified by sequence analysis. RESULTS The amplified products was about 1 375 bp in length, and sequence analysis demonstrated that the N gene fragments had been successfully inserted into pMD18-T vectors. CONCLUSION The successful amplification and cloning of N gene facilitates further investigation of the expression of the N protein and study of its structure and functions.
[Amplification and cloning of the N gene of SARS-associated coronavirus].