CAJ | 학술논문

　　采用 RT-PCR 和 RACE-PCR 技术克隆了大黄鱼(Larimichthys crocea)I 型干扰素基因全长cDNA序列及其基因组序列,并利用荧光定量PCR技术研究了该基因在大黄鱼不同组织中的表达谱,以及副溶血弧菌(Vibrio parahaemolyticus)、LPS、poly I：C刺激后大黄鱼脾脏、肝脏和头肾组织中该基因在转录水平的表达变化。结果表明,大黄鱼I型IFN基因全长cDNA为878bp,开放阅读框558bp,编码185个氨基酸。推测的大黄鱼I型IFN氨基酸序列N端含有20个氨基酸的信号肽,其后包含一个保守的IFabd结构域。大黄鱼I型IFN基因包含5个外显子,4个内含子,在大多数组织和细胞中都有表达,其中在血细胞中表达量最高,肌肉中表达量最低。Poly I：C刺激可诱导大黄鱼I型干扰素基因在脾脏、头肾和肝脏中的表达量显著上调；LPS刺激可诱导大黄鱼I型干扰素基因在脾脏和肝脏中的表达量显著上调；灭活的副溶血弧菌可诱导大黄鱼I型干扰素基因在头肾和肝脏中的表达量显著上调。
　　채용 RT-PCR 화 RACE-PCR 기술극륭료대황어(Larimichthys crocea)I 형간우소기인전장cDNA서렬급기기인조서렬,병이용형광정량PCR기술연구료해기인재대황어불동조직중적표체보,이급부용혈호균(Vibrio parahaemolyticus)、LPS、poly I：C자격후대황어비장、간장화두신조직중해기인재전록수평적표체변화。결과표명,대황어I형IFN기인전장cDNA위878bp,개방열독광558bp,편마185개안기산。추측적대황어I형IFN안기산서렬N단함유20개안기산적신호태,기후포함일개보수적IFabd결구역。대황어I형IFN기인포함5개외현자,4개내함자,재대다수조직화세포중도유표체,기중재혈세포중표체량최고,기육중표체량최저。Poly I：C자격가유도대황어I형간우소기인재비장、두신화간장중적표체량현저상조；LPS자격가유도대황어I형간우소기인재비장화간장중적표체량현저상조；멸활적부용혈호균가유도대황어I형간우소기인재두신화간장중적표체량현저상조。
The full length cDNA and genomic sequence of type I interferon (type I IFN) was cloned from large yellow croaker Larimichthys crocea by RT-PCR and RACE-PCR. The tissue expression and temporal expression profiles of LcIFN I gene after the stimulation with LPS, poly I:C and pathogenic bacteria Vibrio parahemolyticus were described. Our results showed that the full-length cDNA of LcIFN I was 878bp, and open reading frame (ORF) was 558bp, which encoded 185 amino acids. A signal peptide including 20 amino acid residues was at N-terminal, and an IFabd (Interferon alpha, beta and delta) domain was next to signal peptide. LcIFN I genomic sequence consisted of five exons and four introns. Quantitative real-time PCR analysis indicated a broad expression of LcIFN I in most detected tissues, with the most predominant ex-pression in blood cells and the weakest expression in muscle. After injection with poly I:C, LcIFN I expression levels showed up-regulation in head-kidney, liver and spleen; After injection with LPS, LcIFN I expression levels showed up-regulation in liver and spleen; LcIFN I transcripts showed up-regulation in the liver and head-kidney after V. para-hemolyticus injection. Our results suggested that type I IFN might play an important role in large yellow croaker anti-viral and bacterial immune response.