Role of Penaeus monodon Kruppel-like factor (PmKLF) in infection by white spot syndrome virus
Date Issued
2011
Date
2011
Author(s)
Huang, Ping-Han
Advisor
張麗冠
Abstract
Sp1-like proteins and Kruppel-like factors (Sp/KLF) are highly related zincfinger proteins that have crucial roles in transcription. The characteristic feature of this fami- ly is the presence of three zinc fingers in the C-terminal domain, which bind to GC- box, CACCC-box and GT-box sequence, to mediate transcription. This study reports the cloning and characteristics of a Kruppel-like factor (PmKLF) from shrimp, Penae- usmonodon (PmKLF). The full-length PmKLF cDNA is 1702 bps, encoding a poly- peptide of 360 amino acids. Sequence analysis revealed that the sequence of PmKLF is similar to that of KLF11 in humans, mice and zebrafish. Additionally, immunofluo- rescence analysis revealed that GFP-fusion KLF protein is located in the nucleus as dots in an insect cell line, Sf9. Reporter assay revealed that PmKLF enhances the ie1 promoter activity. Moreover, a specific protein-DNA complex binds to the GC-box sequences in the ie1 by electrophoresis mobility shift assays (EMSA). Knockdown of the expression of PmKLF transcript in WSSV-infected shrimp resulted in delayed cu- mulative mortalities, suggesting that PmKLF is important to WSSV infection. Additi- onally, inhibition of PmKLF expression reduced the copy number of WSSV and ie1 expression,revealing that PmKLF affects WSSV infection via interfering with ie1 expression.
Subjects
Kruppel-like factors
GC-box
Penaeus monodon
WSSV
ie1
Type
thesis
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