Degenerate Primed Polymerase Chain Reaction for Detection of Both Nucleopolyhedrovirus and Granulovirus

Document Type : Original Article

Author

Dept of Entomology, Fac of Science, Cairo Univ, Giza, Egypt, 12613.

Abstract

A technique using the polymerase chain reaction
(PCR) was developed for simultaneous detection of the nucleopolyhedrovirus
(NPV) and granulovirus (GV). Ninety one and 73 amino acid sequences of polyhedrin
and granulin genes were compared in pairwise and multiple alignment sequences. Seven
highly conserved DNA sequences within the coding region of the polyhedrin/granulin
genes were identified. Four candidate regions were targeted for amplification
and consequently one pair of degenerate PCR primers was designed to produce
fragments of about 384 bp. The baculoviruses tested by this technique were Autographa
californica (AcMNPV), Bombyx mori NPV (BmNPV), Lymantria
dispar NPV (LdNPV), Spodoptera littoralis NPV (SpliNPV),
S. littoralis GV (SpliGV), Pieris rapae GV (PrGV),
and two local GV isolates (GVG213 and GVF115).
Furthermore, four randomly chosen PCR products were cloned and sequenced. The
sequencing data showed that the four PCR products were fragments of polyhedrin and
granulin genes. Conclusively, this technique would be useful in monitoring the environmental
fate, distribution of Baculovirus species, release of the wild type and
recombinant Baculovirus and quality control studies of Baculovirus
insecticides, as well

Keywords

Egyptian Academic Journal of Biological Sciences. C, Physiology and Molecular Biology
Volume 4, Issue 1
December 2012
Pages 19-23

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