Yabu T, Hirose H, Hirono I, Katagiri T, Aoki T, Yamamoto E
Department of Marine Science and Resources, College of Bioresource Sciences, Nihon University, Kanagawa, Japan.
Mol Mar Biol Biotechnol. 1998 Jun;7(2):138-44.
A complementary DNA library was constructed in lambda ZAP II using messenger RNA from the leukocytes of some heterocloned Japanese flounder, Paralichthys olivaceus, that had been artificially infected with Hirame rhabdovirus (HRV). A cloned flounder interferon regulatory factor (designated fIRF) cDNA was found to be 1746 bp in length, with an open reading frame of 297 amino acids. The overall amino acid sequence of fIRF had approximately 40% identity with the previously reported avian and mammalian IRF-1s and IRF-2s. The fIRF sequence was most similar to that recorded for the chicken IRF-1. Amino acid sequence identities between the DNA-binding domain of the fIRF and that of both chicken IRF-1 and chicken IRF-2 were 72.3%. The DNA-binding domain of fIRF contained the repeated tryptophan motif that is characteristic of members of the IRF family. The mRNA of fIRF was detected in various tissues by reverse transcription-polymerase chain reaction (RT-PCR). The fIRF was transcribed mainly in the intestine, ovary, muscle, liver, heart and spleen, while it was minimally transcribed in the brain and kidney. When Japanese flounder were injected with HRV, the relative expression of fIRF mRNA was found to increase and peak 3 days after injection. The quantities of the fIRF mRNA increased to levels that were 7.5-fold higher than those of noninjected fish. In addition, when Japanese flounder were injected with Edwardsiella tarda, the expression of fIRF mRNA showed increases 2, 3, and 4 days after injection. The quantities of the fIRF mRNA on those days represented approximately 6-, 15-, and 14-fold increases, respectively, over the levels in noninjected fish.
利用人工感染了牙鲆弹状病毒（HRV）的一些异源克隆日本牙鲆（Paralichthys olivaceus）白细胞中的信使核糖核酸，在λZAP II载体中构建了一个互补DNA文库。发现一个克隆的牙鲆干扰素调节因子（命名为fIRF）cDNA长度为1746 bp，具有一个297个氨基酸的开放阅读框。fIRF的整体氨基酸序列与先前报道的禽类和哺乳动物IRF-1及IRF-2约有40%的同一性。fIRF序列与鸡IRF-1的序列最为相似。fIRF的DNA结合结构域与鸡IRF-1和鸡IRF-2的DNA结合结构域之间的氨基酸序列同一性为72.3%。fIRF的DNA结合结构域包含IRF家族成员特有的重复色氨酸基序。通过逆转录-聚合酶链反应（RT-PCR）在各种组织中检测到了fIRF的信使核糖核酸。fIRF主要在肠道、卵巢、肌肉、肝脏、心脏和脾脏中转录，而在脑和肾中的转录最少。当给日本牙鲆注射HRV时，发现fIRF信使核糖核酸的相对表达增加，并在注射后3天达到峰值。fIRF信使核糖核酸的量增加到比未注射鱼高7.5倍的水平。此外，当给日本牙鲆注射迟缓爱德华氏菌时，fIRF信使核糖核酸的表达在注射后2、3和4天显示增加。在这些天，fIRF信使核糖核酸的量分别比未注射鱼的水平增加了约6倍、15倍和14倍。
