[Molecular cloning and sequencing of Dendrolimus punctatus Wenshanensis cytoplasmic polyhedrosis virus (DpwCPV) NS5 protein gene].
DpwCPV were purified from infected Dendrolimus punctatus Wenshanensis larvae by density gradient centrifugation. The genomic dsRNA segments were subsequently extracted directly from the purified polyhedra by SDS treatmen, phenol/chloroform extraction and ethanol precipitation. Defferent segments of genomic dsRNA were separated by low melting point agarose electrophoresis, and S9 dsRNA was purified after being excised from the gel. S9 RNA was denaturalized in high temperature. The primers was designed on the basis of the RNA sequence of S9 in BmCPV-1 (H strain). After RT-PCR, the amplified cDNA was cloned into the pMD18-T Vector. The NS5 protein gene of DpwCPV is found to be 977 nucleotides in length with one open reading frame in one strand. It can code a predicted protein of 320 residues (35560).
Wang
Qiong
Q
Institute of Virology, Wuhan University, Wuhan 430072, China.
Zhang
Jiamin
J
Wen
Li
L
Yang
Juan
J
Hu
Yuanyang
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology*; D012330:RNA, Double-Stranded / Q000235:genetics / Q000302:isolation & purification; D012087:Reoviridae / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics*
D012330:RNA, Double-Stranded; D017361:Viral Nonstructural Proteins