Degenerate Primed Polymerase Chain Reaction for Detection of Both Nucleopolyhedrovirus and Granulovirus | ||
| Egyptian Academic Journal of Biological Sciences. C, Physiology and Molecular Biology | ||
| Article 3, Volume 4, Issue 1, December 2012, Pages 19-23 PDF (192.96 K) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/eajbsc.2012.16121 | ||
| Author | ||
| AlaaEddeen M. Seufi | ||
| 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 | ||
| Nucleopolyhedrovirus; Granulovirus; baculovirus; PCR | ||
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