[The expression and activity detection of a variant N protein of SARS-CoV].Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Jul; 20(4):422-4.XB
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.
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.
(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.
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.