[Amplification and cloning of the N gene of SARS-associated coronavirus].
OBJECTIVE
To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus.


METHODS
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.


CONCLUSIONS
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.
Xiao
Wei-wei
WW
Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China.
Ma
Wen-li
WL
Zhang
Bao
B
Wang
Yan
Y
Mao
Xiang-ming
XM
Peng
Yi-fei
YF
Song
Yan-bin
YB
Wu
Qing-hua
QH
Zheng
Wen-ling
WL
D001483:Base Sequence; D003001:Cloning, Molecular; D000086462:Coronavirus Nucleocapsid Proteins; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics*
D000086462:Coronavirus Nucleocapsid Proteins; D004279:DNA, Viral; D019590:Nucleocapsid Proteins