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ISSN
1672-9145
Acta Biochim Biophys Sin
2004, 36(11): 754–758
CN 31-1940/Q
Expression, Purification and Sublocalization of SARS-CoV
Nucleocapsid Protein in Insect Cells
Ai-Xia REN1,2, You-Hua XIE2, Yu-Ying KONG2,
Guan-Zhen YANG2, Yao-Zhou
ZHANG1,3*, Yuan WANG2*, and Xiang-Fu WU2,3*
1Institute of Biochemistry and Biotechnology,
College of Life Science, Zhejiang University, Hangzhou 310029, China;
2State Key Laboratory of Molecular Biology, Shanghai Institute of
Biochemistry and cell Biology, Shanghai Institutes for Biological Sciences,
Chinese Academy of
Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200031,
China;
3Institute of Biochemistry, Zhejiang University
of Sciences, Hangzhou 310018, China
Abstract The causative agent of
severe acute respiratory syndrome (SARS) is a previously unidentified
coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is a major
viral protein recognized by acute and early convalescent sera from SARS
patients. To facilitate the studies on the function and structure of the N
protein, this report describe the expression and purification of recombinant
SARS-CoV N protein using the baculovirus expression system. Recombinant hexa-histidine-tagged
N protein with a molecular mass of 47 kD was produced in insect cells.
Recombinant N protein was purified to near homogeneity by Ni2+-NTA affinity chromatography. In addition, we examined the
subcellular localization of the N protein by confocal microscopy in Trichoplusia
ni BT1 Tn 5B1-4 cells infected with recombinant baculovirus. The N protein
was found localized in the cytoplasm as well as in the nucleolus. The purified
recombinant N protein can be used in further functional study of SARS-CoV.
Key words severe acute respiratory
syndrome (SARS); SARS coronavirus; nucleocapsid protein; baculovirus; insect
cells
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Received: August 16, 2004
Accepted: October 8, 2004
*Corresponding author: Tel,
86-571-86843190; Fax, 86-571-86843198; E-mail, [email protected]