凡纳滨对虾AP-1基因的克隆和表达特征分析  

Cloning andexpression profile analysis of AP-1 gene from Pacific white shrimp( Litopenaeus vannamei)

作　　者：吴冰[1] 刘逸尘[1] 张亦陈[1] 耿绪云 孙金生[1,2] 

机构地区：[1]天津师范大学生命科学学院,天津市动植物抗性重点实验室,天津300387 [2]天津市水生动物疫病预防控制中心,天津300221

出　　处：《水产学报》2014年第9期1294-1301,共8页Journal of Fisheries of China

基　　金：国家"八六三"高技术研究发展计划(2012AA092205;2012AA10A401);国家"九七三"重点基础研究发展计划(2012CB114405);国家科技支撑计划(2011BAD13B04;2011BAD13B07)

摘　　要：为了探讨凡纳滨对虾转录因子AP-1在病毒引发的免疫应答过程中的潜在作用,实验根据前期的转录组和表达谱结果提示信息,首次克隆了凡纳滨对虾AP-1基因(LvAP-1,GenBank注册号:KF999956),利用在线软件进行了生物信息学分析,运用半定量的方法进行了组织表达分析,并利用实时荧光定量PCR(qPCR)技术分析了该基因在白斑杆状病毒(WSSV)侵染过程中的表达变化特征。结果显示,AP-1基因ORF区全长882 bp,编码293个氨基酸。预测分析显示该基因编码的蛋白质含有1个Jun结构域和1个高度保守的亮氨酸拉链结构域(bZIP),其中Jun结构域在非脊椎动物中保守性不高。组织表达分析表明,该基因在凡纳滨对虾各组织中广泛表达,其中在血细胞中表达量最高。在WSSV感染早期(0.5 hpi),该基因表达没有显著改变,感染后5 h(5 hpi),AP-1基因开始显著上调表达,在人工感染后24 h,该基因的表达量达到最高(P<0.01)。研究表明,该基因在一定程度上参与了凡纳滨对虾体内由WSSV引发的先天免疫应答过程,为进一步研究LvAP-1在对虾应答病毒侵染过程中的功能和作用机制奠定了基础。The purpose is to study Litopenaeus vannamei transcription factor AP-1 potential role in the immune response triggered by the virus. Our previous transcriptome results showed that the expression profile of AP-1 fluctuated significantly in response to WSSV infection. In this research,a novel AP-1 gene（ GenBank Accession Number： KF999956） was cloned from Pacific white shrimp（ L. vannamei）. The full length cDNA of AP-1 has an 882 bp ORF encoding 293 amino acids. The predicted molecular weight and calculated isoelectric point of AP-1 was 32. 7 KDa and 7. 80. The online software（ SMART） analysis results showed that two predicted domains including the Jun protein domain and a highly conserved basic leucine zipper（ bZIP）which existed in the deduced protein of AP-1. Multiple alignment analysis shows that AP-1 shared 48%,42% and 43% amino acid identities with Ixodes Scapularis,Mus musculus and Homo sapiens,respectively.Tissue expression analysis by Semi-quantitative RT-PCR revealed that expression of AP-1 gene is constitutive in all selected tissues and shows highest expression in hemocytes. The transcript of AP-1 is induced in response to white spot syndrome virus（ WSSV） infection. At the early stage of WSSV infection（ 0. 5 hpi）,AP-1 did not change significantly. Then the expression of AP-1 gene began to increase after 5 hpi. At 24 hpi,the gene expression increased significantly and reached the highest level（ P〈0. 01）. From this result,it could be deduced that AP-1 gene took part in shrimp innate immune response caused by WSSV.

关 键 词：凡纳滨对虾 转录因子激活蛋白(AP-1) 序列分析 表达谱分析 免疫应答 

分 类 号：Q785[生物学—分子生物学] S966.1[农业科学—水产养殖]