Selection and expression of peptides which can change the conformation of p20 protein of rice stripe virus.
Phages with high affinity to the P20 protein of rice stripe virus (RSV) were enriched from phage-displayed random 12-mer peptide library after three rounds of phage display screening. Nine different peptides from the enriched library were selected by enzyme-linked immunosorbent assay (ELISA). The P20 protein from raw extracts of rice leaves infected with RSV could be detected by those 9 peptides displayed on the phage, which suggested that a peptide could be an effective tool for diagnosis of RSV in rice and planthopper. Circular dichroism (CD) spectra of P20 fusion proteins with the binding phages and non-binding phages showed that the conformation of P20 protein was changed after binding to each of the 9 selected 12-mer peptides, which suggested that these peptides might disrupt the function of the P20 protein. Thereafter, those peptides might be used to develop plant resistance and disrupt virus transmission. Three of the 12-mer peptide genes were fused with the glutathione-S-transferase (GST) gene in the vector pGEX 3X. The fusion proteins were obtained from an Escherichia coli expression system and purified. The fusion proteins might have a potential to develop a plant peptide-based resistance to its pathogens and virus diagnosis. It also provided a tool (i) to confirm the inhibition of the function of P20 protein by the fusion peptides in vivo, and (ii) to detect the function of P20 protein and the interaction between the virus and its vector.
Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, PR China.
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Zhang
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Xu
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Zhang
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Bai
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Wan
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Shao
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Ye
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Shen
D004591:Electrophoresis, Polyacrylamide Gel; D019151:Peptide Library; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D029364:Tenuivirus / Q000737:chemistry / Q000235:genetics*; D014764:Viral Proteins / Q000737:chemistry*
D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins