Molecular Cloning and Sequencing of Dendrolimus punctatus Wenshanensis Cytoplasmic Polyhedrosis Virus (DpwCPV) NS5 Protein Gene
文山松毛虫质型多角体病毒（DpwCPV）NS5蛋白基因的cDNA克隆及序列分析

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).