Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp

Wang Jing Liu, Yun Shiang Chang, Chung Hsiung Wang, Guang Hsiung Kou, Chu Fang Lo

Research output: Contribution to journalArticlepeer-review

113 Citations (Scopus)

Abstract

Here, we report for the first time the successful use of cycloheximide (CHX) as an inhibitor to block de novo viral protein synthesis during WSSV (white spot syndrome virus) infection. Sixty candidate IE (immediate-early) genes were identified using a global analysis microarray technique. RT-PCR showed that the genes corresponding to ORF126, ORF242 and ORF418 in the Taiwan isolate were consistently CHX-insensitive, and these genes were designated ie1, ie2 and ie3, respectively. The sequences for these IE genes also appear in the two other WSSV isolates that have been sequenced. Three corresponding ORFs were identified in the China WSSV isolate, but only an ORF corresponding to ie1 was predicted in the Thailand isolate. In a promoter activity assay in Sf9 insect cells using EGFP (enhanced green fluorescence protein) as a reporter, ie1 showed very strong promoter activity, producing higher EGFP signals than the insect Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV) ie2 promoter.

Original languageEnglish
Pages (from-to)327-341
Number of pages15
JournalVirology
Volume334
Issue number2
DOIs
Publication statusPublished - 2005 Apr 10

All Science Journal Classification (ASJC) codes

  • Virology

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