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Virologica Sinica, 18 (1) : 39-43, 2003
M olecular Cloning Dendrolimus punctatus cytoplasmic polyhedrosis virus Segment 8 by Single Primer Amplification and Sequence Analysis
Institutl of,Virology,Wuhan University,Wuhan 430072,China
Dendrolimus punctatus cytoplasmic polyhedrosis virus(DpCPV)was purified from infected
Dendrolimus punctattts and the genomic dsRNA segments were subsequently extracted. Segment 8
dsRNA was purified by low melting-temperat agraose.A single amino-linked modified oligonucleotide
(primer 1) was ligated to either 3’end of dsRNA genome segment 8 by Using T4 RNA Ligase.
Following reverse transcription, annealing and repair of cDNA strand. amplification of S8 dsRNA
genome was accomplished by PCR using a single complementary oligonucleotidc(primer 2).The
amplified cDNA was cloned into the pMD18-T vector.Nucleotide sequence analysis of cDNA derived
from the segment 8 revealed that it consists of 1 330 nucleotides encoding putative protein of 390 amino
acids with molecular weith of 44KD. Comparison of the nucleolide sequence of the segment 8 of
DpCPV with that of BmCPV and LdCPV showed that nucleotides homology is 83% and 97% .
  Published online: 2 Feb 2003
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