Molecular analysis of an occlusion body protein from Culex nigripalpus nucleopolyhedrovirus (CuniNPV).
CUN085 is an occlusion body (OB) protein from the nucleopolyhedrovirus of Culex nigripalpus (CuniNPV). SDS-PAGE analysis indicated that the CuniNPV OB protein is about 3 times the size (approximately 90 kDa) of characterized nucleopolyhedrovirus (NPVs) and granulovirus OB proteins. Rapid amplification of cDNA ends (RACE), RNase protection assay, real-time PCR, and protein sequencing were used to characterize CUN085 from CuniNPV. RACE data revealed that the transcriptional start and termination sites for the CUN085 gene yielded a polypeptide comprised of 822 amino acids indicating that translation initiates within a larger 882 amino acid open reading frame that was originally predicted from the CuniNPV genome sequence. Transcription of CUN085 started at a consensus baculovirus late transcription start site TAAG at nucleotide position 75433 of the CuniNPV genome sequence. RNase protection assays and quantitative real-time PCR show that the CUN085 transcript is first detected in mosquito larvae at approximately 6 h after infection with CuniNPV and its prevalence increased progressively over the subsequent 18 h.
Perera
Omaththage P
OP
Southern Insect Management Research Unit, USDA-ARS, Stoneville, MS 38776, USA. operera@msa-stoneville.ars.usda.gov
Valles
Steven M
SM
Green
Terry B
TB
White
Susan
S
Strong
Charles A
CA
Becnel
James J
JJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003465:Culex / Q000821:virology*; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics*
D014764:Viral Proteins