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PubMed Central
Article . 2004
Data sources: PubMed Central
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Current Medical Science
Article . 2004 . Peer-reviewed
License: Springer Nature TDM
Data sources: Crossref
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Expression and purification of SARS coronavirus membrane protein

Authors: Wuxing, Dai; Mingjun, Lei; Shaoting, Wu; Zhihao, Chen; Liang, Liang; Hujrong, Pan; Li, Qin; +3 Authors

Expression and purification of SARS coronavirus membrane protein

Abstract

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.

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Keywords

Reverse Transcriptase Polymerase Chain Reaction, Membrane Proteins, Viral Vaccines, Article, Recombinant Proteins, Severe acute respiratory syndrome-related coronavirus, Escherichia coli, Humans, Cloning, Molecular, Plasmids

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average
Green
bronze