[Over-expression in Escherichia coli and purification of nucleocaspid and membrane protein of SARS coronavirus]

Sheng Wu Gong Cheng Xue Bao. 2003 Jul;19(4):392-6.
[Article in Chinese]

Abstract

Genes encoding nucleocaspid (N) and membrane (M) protein of SARS coronavirus were obtained by RT-PCR and were cloned into expression vector pET22b and pBV222. DNA sequencing showed that the genes cloned from a patient in Beijing were identical to the gene sequences from reported Toronto strain. The genes were over-expressed in E. coli either as inclusion body or as soluble form. The recombinant proteins were purified by ion-exchange, or ion-exchange followed by metal chelate affinity chromatography. The recombinant N protein was demonstrated highly antigenic and could be employed as antigen to detect SARS antibodies in ELISA system for SARS diagnosis.

MeSH terms

  • Chromatography, Affinity
  • Chromatography, Ion Exchange
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Nucleocapsid Proteins / genetics
  • Nucleocapsid Proteins / isolation & purification*
  • Nucleocapsid Proteins / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Severe acute respiratory syndrome-related coronavirus / genetics
  • Severe acute respiratory syndrome-related coronavirus / metabolism*
  • Viral Structural Proteins / genetics
  • Viral Structural Proteins / isolation & purification*
  • Viral Structural Proteins / metabolism*

Substances

  • Nucleocapsid Proteins
  • Viral Structural Proteins