凡纳滨对虾亲环素A基因的克隆及表达分析

凡纳滨对虾亲环素A基因的克隆及表达分析 Cloning and Expression Analysis of Cyclophilins A Gene from Litopenaeus vannamei 丁兆坤 DING Zhaokun1 1 first-author 辛文伦 XIN Wenlun1 1 陈秀荔 CHEN Xiuli2 2 赵永贞 ZHAO Yongzhen2 2 许友卿 XU Youqing1 1 陈晓汉 CHEN Xiaohan2 2 广西大学水产科学研究所 广西大学水产科学研究所 Institute for Fishery Sciences,Guangxi University Institute for Fishery Sciences,Guangxi University 广西水产研究所广西水产遗传育种与健康养殖重点实验室 广西水产研究所广西水产遗传育种与健康养殖重点实验室 Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Aquaculture,Guangxi Instituteof Fisheries Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Aquaculture,Guangxi Instituteof Fisheries 通过电子克隆所得基因序列设计引物,采用RT-PCR方法首次克隆得到长度为885 bp的凡纳滨对虾亲环素A（Cyclophilins A）CYPA基因cDNA序列,其开放阅读框为35 5 29 bp,可编码164个氨基酸.推算其分子量约为17.6×103,理论等电点为8.55.与其他物种的CYPA基因比对发现,该基因的氨基酸序列具有较高的保守性.基于氨基酸序列的聚类分析表明,凡纳滨对虾与斑节对虾的亲缘关系最近.荧光定量PCR的结果显示该基因在组织中广谱表达,在抗病组中,胃中的表达量最高,心脏中的表达量最低,差异约为2倍.而在感病组中,该基因在心脏中的表达量最高,肌肉中的表达量最低.利用ProtFun在线预测蛋白质的功能分类,表明CYPA基因可能是一免疫应答抑制因子,参与凡纳滨对虾免疫防御反应. Cyclophilins A cDNA gene（CYPA gene） with 885 bp was firstly cloned from Litopenaeus vannamei by RT-PCR.Its open reading frame（ORF） was 35-529 bp,coding 164 amino acids.The predicted molecular weight was about 17.6×103,and its calculated isoelectric point（pI） was 8.55.The sequence of CYPA gene was compared with that of the other species through BLAST analysis,indicating that the amino acid sequence of CYPA gene cloned from Litopenaeus vannamei was highly conserved.Based on the amino acid sequences of the gene,phylogenetic tree showed that Litopenaeus vannamei was closely related to Penaeus monodon.Broad-spectrum expression was found in the CYPA gene from the tissues of Litopenaeus vannamei.However,in the antiviral population,the CYPA gene expression was the highest in the stomach and the lowest in the heart,in which the difference was about 2 folds.While in the viral-challenge population,the CYPA gene expression was the highest in the heart and the lowest in the muscle.According to the ProtFun online prediction of protein functional classification,the result demonstrated that the CYPA gene might be an immune response inhibitor participated in the immune defense response in Litopenaeus vannamei. 凡纳滨对虾 亲环素A 克隆 荧光定量PCR Litopenaeus vannamei cyclophilin A clone RT-PCR 10.14188/j.1671-8836.2011.04.006 S917.4 农业部现代农业产业技术体系建设专项资金（CARS-47）;广西壮族自治区直属公益性科研院所基本科研业务费专项资金项目（2060302CXIF-2009-02）;广西壮族自治区自然科学基金项目（桂科基0836028桂科自0832009Z） 2011-04-01 2021-04-01 4 57