乌鬃鹅RIG-Ⅰ基因的克隆及初步分析.pdfVIP

⑨怕在舍2014 年第36 卷第 19 期 China Poultry Vol.36.No.19.2014 。遗传育种。 乌番争鹅RIG- 1 基因的克隆及初步分析* 吴宁昭，王彬，左晓昕，陈卓，张阿秀，陈阳，徐琪，陈国宏 (扬州大学动物科学与技术学院，江苏扬州 225009) 摘 要:维甲酸诱导基因蛋白 1 (RIG- 1)是一类胞质内模式识别受体，在机体抗病毒天然 免疫过程中发挥重要作用。本研究通过poly(I :C)和5ppp-dsRNA剌激原代鹅胚成纤维细胞， 采用 RT-PCR扩增获得了乌警鹅RIG- 1 CDS，其全长2805 bp，编码934个氨基酸。氨基酸序 列分析表明，该蛋白无跨膜区，无信号肤，位于细胞质和细胞核，故推测gRIG- 1 蛋白为胞质内 可溶性蛋白。实时荧光定量PCR 结果显示，poly (I: C)和5ppp-dsRNA 能显著上调gRIG- 1 mRNA水平。研究结果为RIG- 1 在鹅抗病毒天然免疫研究奠定了一定的理论基础。 关键词:鹅;维甲酸诱导基因蛋白 1 ;基因克隆;反转录聚合酶链反应 Cloning and Expression Analysis of RIG- 1 Gene in Wuzong Goose. WU Ningzhao ,WANG Bin ,ZUO Xiaoxin ,CHEN Zhuo ,ZHANG Axiu , CHEN Yang ,XU Qi ,CHEN Guohong (College of Animal Science and Technology ,Yangzhou University ,Yangzhou ,Jiangsu 225009) Abstract: Retinoic acid inducible gene- 1 (RIG- 1 )is a kind of protein that acts as an intracellular pattern recognition receptors , which plays an important role in the immune response. In this study , RIG- 1 coding sequence ( CDS) of Wuzong goose was amplified by RT-PCR under the stimulation of poly (1: C) and 5 triphosphate double stranded RNA (5 ppp- dsRNA) by separating the original generation of goose embryonic fibroblasts. The sequencing results showed that full-length sequence of goose RlG- 1 gene CDS was 2 805 bp , encoding 934 amino acids. AA sequence analysis showed that there was no signal peptide sequence and no transmembrane domain for gRIG. Subcellular localization analysis predicted that RIG- 1 would be mostly located in the cytoplasm and nucleus. Therefore it was deduced that gRIG- 1 was an intracellular hydrophilic protein. Real- time PCR results showed that gRIG- 1 mRNA e