Department of Biochemical Science and Technology, National Taiwan University, Taipei 106, Taiwan, ROC.
Dev Comp Immunol. 2012 Jan;36(1):121-9. doi: 10.1016/j.dci.2011.06.008. Epub 2011 Jun 28.
Sp1-like proteins and Kruppel-like factors (KLFs) are highly related zinc-finger proteins that have crucial roles in transcription. One expressed sequence tag (EST, HPA-N-S01-EST0038) from shrimps is homologous to Sp1. This study reports the cloning and characteristics of a KLF from shrimp, Penaeus monodon (PmKLF). The full-length PmKLF cDNA is 1702 bp, encoding a polypeptide of 360 amino acids. Sequence analysis revealed that the sequence of PmKLF is similar to that of KLF11 in humans, mice and zebrafish. RT-PCR analysis indicated that PmKLF mRNA is expressed in all examined tissues. Additionally, immunofluorescence analysis revealed that GFP-KLF fusion protein is located in the nucleus as dots in an insect cell line, Sf9. Localization of PmKLF in the nucleus is also observed in the hemolymph from white spot syndrome virus (WSSV)-infected and WSSV-uninfected Litopenaeus vannamei. Knockdown of the expression of PmKLF transcript in WSSV-infected shrimp resulted in delayed cumulative mortalities, suggesting that PmKLF is important to WSSV infection. Moreover, inhibition of PmKLF expression reduced the copy number of WSSV and ie1 expression, revealing that PmKLF affects WSSV infection via interfering with ie1 expression.
Sp1 样蛋白和 Kruppel 样因子（KLFs）是高度相关的锌指蛋白，在转录中具有关键作用。从虾中获得的一个表达序列标签（EST，HPA-N-S01-EST0038）与 Sp1 同源。本研究报告了一种来自虾类（Penaeus monodon）的 KLF 的克隆和特性（PmKLF）。PmKLF 的全长 cDNA 为 1702bp，编码 360 个氨基酸的多肽。序列分析表明，PmKLF 的序列与人、小鼠和斑马鱼的 KLF11 相似。RT-PCR 分析表明，PmKLF mRNA 在所有检测的组织中均有表达。此外，免疫荧光分析显示，GFP-KLF 融合蛋白在昆虫细胞系 Sf9 中呈点状存在于细胞核内。在感染和未感染白斑综合征病毒（WSSV）的凡纳滨对虾的血淋巴中也观察到 PmKLF 的核定位。在感染 WSSV 的虾中敲低 PmKLF 转录本的表达导致累积死亡率延迟，表明 PmKLF 对 WSSV 感染很重要。此外，抑制 PmKLF 的表达降低了 WSSV 和 ie1 表达的拷贝数，表明 PmKLF 通过干扰 ie1 表达影响 WSSV 感染。
