Selection of phage-display peptides that bind specifically to the outer coat protein of Rice black streaked dwarf virus.
Several peptides that could bind specifically to the outer coat protein encoded by the S10 gene of Rice black streaked virus (RBSDV) were isolated from a phage-display random 12-mer peptide library. The sequence analysis showed that the amino acid motif (K)K**(*)P, the asterisk denoting any amino acid, might be the core sequence by which the peptides bind to the target protein. The peptide 1 that had a high affinity to RBSDV outer coat protein was synthesized by a chemical method and its fusion protein with glutathione-S-transferase (GST) was produced in an Escherichia coli expression system. The dot and Western blot analyses indicated that RBSDV could be detected with a high sensitivity in crude extracts of diseased plant leaves using a purified GST fusion protein. The circular dichroism (CD) spectroscopy revealed that the synthesized binding peptide but not a nonbinding peptide could bring about a marked change in the conformation of outer coat RBSDV protein. Since the protein functions only when it has correct conformation, the peptides binding specifically to it could possibly disturb the function of the virus outer coat protein and might be used to block the transmission pathway of the virus. Summing up, as these peptides showed a high specificity and sensitivity and diagnostic potential for RBSDV, they may represent the basis of a novel strategy for development of resistance to RBSDV.
Bai
F W
FW
Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, PR China.
Zhang
H W
HW
Yan
J
J
Qu
Z C
ZC
Xu
J
J
Wen
J G
JG
Ye
M M
MM
Shen
D L
DL
D019677:Artificial Gene Fusion; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D036022:Capsid Proteins / Q000302:isolation & purification / Q000378:metabolism*; D015870:Gene Expression; D005982:Glutathione Transferase / Q000378:metabolism; D008969:Molecular Sequence Data; D012275:Oryza / Q000821:virology; D019151:Peptide Library; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism; D010942:Plant Viruses / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D012087:Reoviridae / Q000235:genetics* / Q000378:metabolism; D012680:Sensitivity and Specificity
D036022:Capsid Proteins; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase