Molecular characterization of avian reoviruses using nested PCR and nucleotide sequence analysis.
A nested polymerase chain reaction (PCR) with subsequent nucleotide sequence analysis identified and differentiated avian reoviruses (ARVs). PCR products amplified from the S1 gene segment of ARV of USA isolates were 738 and 342 bp, respectively. PCR products were conformed by Southern and dot blot hybridizations. The amplified cDNA fragments were cloned into the pUC18 vector and subjected to DNA sequencing. The nucleotide and deduced amino acid sequences of four USA (S1133, 1733, 2408, and CO8) and two Australian isolates (RAM-1 and SOM-4) were compared. Results of paired difference analysis and a predicted dendrogram revealed that USA isolates were closely related, but different from, Australian isolates. The deduced amino acid sequences of the N-terminal region of ARV sigma C showed a heptapeptide repeat of hydrophobic residues in all ARV isolates.
Liu
H J
HJ
Department of Poultry Science, Auburn University, AL 36849-5416, USA.
Giambrone
J J
JJ
Nielsen
B L
BL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002642:Chick Embryo; D003001:Cloning, Molecular; D005347:Fibroblasts; D015151:Immunoblotting; D008969:Molecular Sequence Data; D018165:Orthoreovirus / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014764:Viral Proteins / Q000235:genetics
D014764:Viral Proteins