【摘要】 FcβGBP-HDL是中国明对虾免疫应答中的一种模式识别受体,在白斑综合征病毒(white spot syndrome virus,WSSV)侵染后其表达显著上调。为了探讨FcβGBP-HDL在免疫应答中的转录调节机制,实验以FcβGBP-HDL cDNA为基础设计引物,采用染色体步移技术对其基因组DNA和启动子进行克隆,构建一系列缺失表达载体进行启动子活性分析。结果显示,FcβGBP-HDL基因全长6 713 bp,无内含子;启动子区域长1 507 bp,除了1个启动子核心序列、2个SRF、2个TBP、1个CTF和1个CRE等典型的启动子特征外,还有2个GATA-1、3个AP-1、1个c-Ets-1和7个Sp1等参与其它节肢动物免疫基因转录调节的结合位点;同源比对分析表明,FcβGBP-HDL启动子与FcCTL启动子相似度最高(41.4%);不同长度的启动子片段具有不同的启动荧光素酶报告基因表达的活性,其中去除5’端2个AP-1后的启动子片段具有最高的启动活性,去除启动子核心序列的启动子片段启动活性最低。研究表明,FcβGBP-HDL启动子可能在免疫反应中受到调节。更多还原

【Abstract】 FcβGBP-HDL was a pattern recognition receptor(PRR)which was up-regulated post WSSV(white spot syndrome virus)challenge.To investigate the mechanism of transcription regulation,primers were designed based on the FcβGBP-HDL cDNA obtained,the gene and 5′-flanking sequences were isolated from the Chinese Shrimp Fenneropenaeus chinensis by genome walking,and the activity of promoter was analyzed by voter construction.The results showed that this gene was composed of only one exon spanning 6 713 bp.The 5′-flanking sequence(1 507 bp)of FcβGBP-HDL gene contained several typical transcription regulation elements,such as one promoter core sequence,two SRF,two TBP,one CTF and one CRE,and several transcription regulation elements of immune-gene in other arthropods,such as two GATA-1,three AP-1 and seven Sp1.The upstream sequence of FcβGBP-HDL gene had highest level similarity with FcCTL(similarity 41.4%).The transcription levels of luciferase reporter gene were up-regulated in all construction voters,highest in pGL3-FcβGBP2 with the deletion of two AP-1,and lowest in pGL3-FcβGBP4 with the deletion of promoter core sequence.The results indicated that promoter of FcβGBP-HDL might be regulated in immune reaction.更多还原