[The expression and activity detection of a variant N protein of SARS-CoV].
OBJECTIVE
To construct an expression vector pGEX-2T/N, and to express the fusion protein consisting of N protein of SARS-CoV and GST in E.coli.


METHODS
The N region gene of SARS-CoV was cloned by RT-PCR. The expression vector was constructed by DNA recombination. The recombinant plasmid was transformed into E.coli BL21(DE3). The expression of the fusion protein was detected by Western blot.


RESULTS
(1) As compared with the sequences in GenBank, 20 bp were deleted in DNA sequence of the cloned N protein. (2) The fusion protein GST-N was soluble. Western blot analysis showed that the reaction of GST-N to anti-SARS-CoV sera was positive.


CONCLUSIONS
The pGEX-2T/N has been constructed and expressed in the form of fusion protein GST-N successfully, which lays the foundation for further study of SARS-CoV N protein.
Qi
Yang
Y
Department of Immunology, Institute of Infectious Diseases, 302th Hospital of PLA, Beijing 100039, China. qiy2004@sohu.com
Zheng
Yu
Y
Shu
Cui-li
CL
Jiang
Li
L
Hu
Yan
Y
Mao
Pan-yong
PY
Cheng
Yun
Y
D001483:Base Sequence; D000086462:Coronavirus Nucleocapsid Proteins; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics*; D006801:Humans; D019590:Nucleocapsid Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics*; D017384:Sequence Deletion; D014162:Transfection
D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase