CAJ | 학술논문

IFN-γ在天然免疫和适应性免疫应答中均发挥着重要的作用.由于低等脊椎动物 IFN-γ与哺乳动物 IFN-γ相似性非常低,硬骨鱼类的 IFN-γ近年来才得到鉴定.本研究报道了斜带石斑鱼(Epinephelus coioides)的 IFN-γ基因,并对其表达进行了分析.斜带石斑鱼 IFN-γ cDNA 全长为867 bp,编码200个氨基酸.其基因组全长为5104 bp,由4个外显子、3个内含子组成.氨基酸序列比对显示斜带石斑鱼 IFN-γ与高等脊椎动物 IFN-γ的序列相似性较低,仅为32.5%~39%,与其他硬骨鱼类的 IFN-γ相似性为38.0%~68.5%.序列分析结果显示,斜带石斑鱼 IFN-γ分子 C末端中存在 IFN-γ的特征序列以及核定位基序.二级结构分析结果显示斜带石斑鱼 IFN-γ序列中包含6个α螺旋结构,其排列与高等脊椎动物的 IFN-γ类似.应用荧光定量 PCR 检测了 IFN-γ基因在 Poly I:C 诱导后的表达变化.发现在肾、脾、鳃、头肾、皮肤和胸腺组织中, Poly I:C 能显著诱导 IFN-γ的表达,在胸腺中上调倍数最高,其次为头肾、脾、肾、皮肤和鳃,而在脑和心脏组织中没有显著变化.据此推测, IFN-γ在斜带石斑鱼抵抗病毒感染的过程中起有十分重要的作用.
IFN-γ재천연면역화괄응성면역응답중균발휘착중요적작용.유우저등척추동물 IFN-γ여포유동물 IFN-γ상사성비상저,경골어류적 IFN-γ근년래재득도감정.본연구보도료사대석반어(Epinephelus coioides)적 IFN-γ기인,병대기표체진행료분석.사대석반어 IFN-γ cDNA 전장위867 bp,편마200개안기산.기기인조전장위5104 bp,유4개외현자、3개내함자조성.안기산서렬비대현시사대석반어 IFN-γ여고등척추동물 IFN-γ적서렬상사성교저,부위32.5%~39%,여기타경골어류적 IFN-γ상사성위38.0%~68.5%.서렬분석결과현시,사대석반어 IFN-γ분자 C말단중존재 IFN-γ적특정서렬이급핵정위기서.이급결구분석결과현시사대석반어 IFN-γ서렬중포함6개α라선결구,기배렬여고등척추동물적 IFN-γ유사.응용형광정량 PCR 검측료 IFN-γ기인재 Poly I:C 유도후적표체변화.발현재신、비、새、두신、피부화흉선조직중, Poly I:C 능현저유도 IFN-γ적표체,재흉선중상조배수최고,기차위두신、비、신、피부화새,이재뇌화심장조직중몰유현저변화.거차추측, IFN-γ재사대석반어저항병독감염적과정중기유십분중요적작용.
@@@@IFN-γ is a typeⅡinterferon that plays an important role in both innate and cell-mediated immune re-sponses. Because of the low similarity with their mammalian homologues, IFN-γ in teleost fish has only recently been identified. In this study, the cDNA sequence encoding the IFN-γ molecule was cloned from the or-ange-spotted grouper, Epinephelus coioides, and its gene organization was revealed. Its expression in response to Poly I:C stimulation was also examined. The IFN-γ cDNA sequence consists of 867 bp, which encodes a putative protein of 200 amino acids. The gene is 5-kb long and contains four exons, which are comparable in size with IFN-γ genes in other vertebrates. This IFN-γ was only 32.5%?39% similar to its counterparts in higher vertebrates and 38.0%?68.5% similar to those in other teleost fish. Sequence analysis revealed that the IFN-γ in the or-ange-spotted grouper contains the typical IFN-γ motif and a conserved nuclear localization site in the C-terminal end and displays an alpha-helix structure similar to mammalian IFN-γ. Real time PCR analysis results showed that the expression of IFN-γ increased in the thymus, head kidney, spleen, kidney, skin, and gills following the poly I:C stimulation, indicating its role against viruses in orange-spotted grouper.