TY - JOUR
T1 - Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp
AU - Liu, Wang Jing
AU - Chang, Yun Shiang
AU - Wang, Chung Hsiung
AU - Kou, Guang Hsiung
AU - Lo, Chu Fang
N1 - Funding Information:
This investigation was supported financially by the National Science Council grants (NSC93-2311-B-002-013, NSC93-2317-B-002-017, and NSC93-2317-B-002-018). Both Guang-Hsiung Kou and Chu-Fang Lo are corresponding authors for this work. We are indebted to Mr. Paul Barlow for his helpful criticism of the manuscript.
PY - 2005/4/10
Y1 - 2005/4/10
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=15244343329&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=15244343329&partnerID=8YFLogxK
U2 - 10.1016/j.virol.2005.01.047
DO - 10.1016/j.virol.2005.01.047
M3 - Article
C2 - 15780883
AN - SCOPUS:15244343329
SN - 0042-6822
VL - 334
SP - 327
EP - 341
JO - Virology
JF - Virology
IS - 2
ER -