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Transcriptome sequencing of and microarray development for a Helicoverpa zea cell line to investigate in vitro insect cell-baculovirus interactions

ABSTRACT: The Helicoverpa armigera single-capsid nucleopolyhedrovirus (HaSNPV) can be propagated using H. zea insect cell cultures, for use as a biopesticide against Heliothine agricultural pests.This study sequenced, assembled and functionally annotated 29,586 transcript sequences from cultured H. zea cells using Illumina 100 bps and paired-end transcriptome sequencing (RNA-seq). From these sequences, a genome-scale microarray platform was constructed and validated for effective expression analysis of H. zea genes. This array also included probes for all HaSNPV genes, thereby allowing virus and host gene changes to be monitored simultaneously. A 4x180,000 SurePrint Agilent expression array (Agilent, Santa Clara, CA) was employed so that a high number of probes can be included to test all 29,586 assembled H. zea sequences and to eventually select a best probe for each transcript. Two biological replicates for uninfected H. zea cells and two other biological replicates for infected samples at 18 hours post infection were analyzed. Six Agilent 60-mer oligonucleotide probes were designed by eArray (Agilent) for each transcript, in which each orientation had three probes randomly distributed across the sequences. Probes that had potential cross-hybridization (Xhyb) were removed. The final probe set for H. zea sequences included 153,583 probes. In addition, probes for all 135 H.armigera single-capsid nucleopolyhedrovirus (HaSNPV) genes were added to investigate host-virus interaction in culture.

ORGANISM(S): Helicoverpa zea

SUBMITTER: Quan Nguyen

PROVIDER: E-GEOD-34418 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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