Selection of a specific peptide from a nona-peptide library for in vitro inhibition of grass carp hemorrhage virus replication.
Grass carp hemorrhage virus (GCHV), a member of Reoviridae, causes severe hemorrhagic disease of grass carp (Ctenopharyngodon idellus) in China. Icosahedral virions of GCHV were used as to assay the effect of specific peptides for the inhibition of GCHV infectivity. A random nona-peptide library displayed on phage fUSE5 was constructed, and the expressed peptides were fused onto the amino terminus of the minor coat protein III. By biopanning, the fused peptides were bound to the biotinylated GCHV. Phages containing specific peptides bound to GCHV were eluted and amplified in Escherichia coli K91. Three rounds of affinity selection enriched the pool of inhibiting peptides. Sixteen clones which inhibited the replication of GCHV in a grass carp kidney cell line were selected. The TCID(50) of GCHV was decreased over 10000x. Six clones having the strongest inhibitory effect shared the same DNA sequence, with a deduced amino acid sequence of NH(2)-Leu-Trp-Val-Gly-Gly-Gly-Arg-Asn-Ala-COOH. A synthesized nona-peptide of identical sequence exhibited similar inhibitory activity towards GCHV replication in vitro.
Wang
B
B
Institute of Microbiology, Chinese Academy of Sciences, Beijing, PR China.
Ke
L H
LH
Jiang
H
H
Li
C Z
CZ
Tien
P
P
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000494:pharmacology; D002347:Carps / Q000821:virology*; D002460:Cell Line; D002681:China; D003001:Cloning, Molecular; D003588:Cytopathogenic Effect, Viral; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000494:pharmacology*; D012087:Reoviridae / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D019151:Peptide Library; D010455:Peptides