Xiao Wei-wei, Ma Wen-li, Zhang Bao, Wang Yan, Mao Xiang-ming, Peng Yi-fei, Song Yan-bin, Wu Qing-hua, Zheng Wen-ling
Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2004 Jan;24(1):39-41.
To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus.
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.
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.
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.