High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells

Fei Zhong, Zhen Yu Zhong, Shuang Liang, Xiu Jin Li

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Aim: To develop a highly efficacious method for preparation of soluble SARS S-protein using adenovirus vector to meet the requirement for S-protein investigation. Methods: The human adenovirus vector was used to express the soluble S-protein (corresponding to 1-1190 amino acids) fused with Myc/His tag using codon-optimized gene construct in HEK239 cells. The recombinant adenovirus bearing S-protein gene was generated by ligation method. The expressed S-protein with Myc/His tag was purified from culture medium with Ni-NTA agarose beads followed by dialysis. The S-protein was detected by Western blot and its biologic activity was analyzed by binding to Vero cells. Results: Under the conditions of infection dose (MOI of 50) and expression time (48 h), the high-level expression of S-protein was obtained. The expression level was determined to be approximately 75 μg/10 6 cells after purification. Purified soluble S-protein was readily detected by Western blot with anti-Myc antibody and showed the ability to bind to surface of Vero cells, demonstrating that the soluble S-protein could remain the biologic activity in the native molecule. Conclusion: The high-level expression of S-protein in HEK293 cells mediated by adenovirus can be achieved under the optimized expression conditions. The proteins possess the biologic activity, which lays a foundation for further investigation of S-protein biological function.

Original languageEnglish (US)
Pages (from-to)1452-1457
Number of pages6
JournalWorld Journal of Gastroenterology
Volume12
Issue number9
DOIs
StatePublished - Mar 7 2006
Externally publishedYes

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HEK293 Cells
Protein S
Adenoviridae
Proteins
Vero Cells
Western Blotting
Human Adenoviruses
Codon
Sepharose
Ligation
Culture Media
Dialysis
Anti-Idiotypic Antibodies
Amino Acids

Keywords

  • Expression
  • S-protein
  • SARS-CoV

ASJC Scopus subject areas

  • Gastroenterology

Cite this

High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells. / Zhong, Fei; Zhong, Zhen Yu; Liang, Shuang; Li, Xiu Jin.

In: World Journal of Gastroenterology, Vol. 12, No. 9, 07.03.2006, p. 1452-1457.

Research output: Contribution to journalArticle

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