Dissection of SARS Coronavirus Spike Protein into Discrete Folded Fragments |
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Authors: | LI Shuang CAI Zhen CHEN Yong LIN Zhanglin |
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Affiliation: | Laboratory of Directed Evolution, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China |
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Abstract: | The spike protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) mediates cell fusion by binding to target cell surface receptors. This paper reports a simple method for dissecting the viral protein and for searching for foldable fragments in a random but systematic manner. The method in-volves digestion by DNase I to generate a pool of short DNA segments, followed by an additional step of re-assembly of these segments to produce a library of DNA fragments with random ends but controllable lengths. To rapidly screen for discrete folded polypeptide fragments, the reassembled gene fragments were further cloned into a vector as N-terminal fusions to a folding reporter gene which was a variant of green fluorescent protein. Two foldable fragments were identified for the SARS-CoV spike protein, which coincide with various anti-SARS peptides derived from the hepated repeat (HR) region 2 of the spike protein. The method should be applicable to other viral proteins to isolate antigen or vaccine candidates, thus providing an alternative to the full-length proteins (subunits) or linear short peptides. |
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Keywords: | severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein dissection foldable fragment green fluorscent protein (GFP) |
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