Premium
Comparison of three methods of parasitoid polydnavirus genomic DNA isolation to facilitate polydnavirus genomic sequencing
Author(s) -
RodríguezPérez Mario A.,
Beckage Nancy E.
Publication year - 2008
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.20228
Subject(s) - biology , genome , genetics , genomic dna , gene , dna sequencing , diatraea saccharalis , virology , botany , pest analysis
A major long‐term goal of polydnavirus (PDV) genome research is to identify novel virally encoded molecules that may serve as biopesticides to target insect pests that threaten agriculture and human health. As PDV viral replication in cell culture in vitro has not yet been achieved, several thousands of wasps must be dissected to yield enough viral DNA from the adult ovaries to carry out PDV genomic sequencing. This study compares three methods of PDV genomic DNA isolation for the PDV of Cotesia flavipes , which parasitizes the sugarcane borer, Diatraea saccharalis , preparatory to sequencing the C. flavipes bracovirus genome. Two of these protocols incorporate phenol‐chloroform DNA extraction steps in the procedure and the third protocol uses a modified Qiagen DNA kit method to extract viral DNA. The latter method proved significantly less time‐consuming and more cost‐effective. Efforts are currently underway to bioengineer insect pathogenic viruses with PDV genes, so that their gene products will enhance baculovirus virulence for agricultural insect pests, either via suppression of the immune system of the host or by PDV‐mediated induction of its developmental arrest. Sequencing a growing number of complete PDV genomes will enhance those efforts, which will be facilitated by the study reported here. Arch. Insect Biochem. Physiol. 67:202–209, 2008. © 2008 Wiley‐Liss, Inc.