IHHNV病毒蛋白与凡纳滨对虾围食蛋白的互作研究-水产养殖专业毕业论文.docx

之间的作用较弱，怀疑其为假阳性。由此可见，LvPT蛋白与IHHNV病毒 间产生相互作用，而且有可能直接影响病毒侵袭机体的过程。 此外，我们采用RNAi的方法，体外合成dsLvPT后，进行体内注射干 扰LvPT基因的表达，研究IHHNV病毒在凡纳滨对虾体内复制是否受到抑 制。结果表明，注射dsLvPT后，LvPT表达下调，IHHNV病毒的相对拷贝 数与对照组相比显著降低。进一步说明LvPT与IHHNV病毒感染凡纳滨对 虾的机制有关。此外，我们通过western-blot检测发现LvPT可以在High-Five 昆虫细胞中表达，为进一步研究LvPT与IHHNV病毒编码的蛋白间的相互 作用机制奠定了基础。 关键词：凡纳滨对虾 IHHNV 围食膜蛋白RNAi 万方数据  INTERACTIoN RESEARCH of IHIINV PRoTEINS AND LIToPENAEUS Vf心NAMEI PERITRoPHIC PRoTEINS ABSTRACT The intestinal tissue eDNA library of Litopenaeus vannamei was constructed successfully by using SMART technology．A full—length sequence of peritrophic in Litopenaeus vannamei was obtained by electronic splicing and sequence analysis．The sequence encodes 275 amino acids，with a theoretical molecular weight of about 30．782 kDa．Comparative analysis of the Peritrophin gene sequence obtained in the further study showed the similarities of 77．3％， 76．1％，72．2％，and 62．3％compared to the sequences of Penaeus monodon， Penaeus semisulcatus，Chinese shrimp，and Penaeus merguiensis．According to the comparison results，a conservative segment ofL．vannamei Peritrophin(LvPT) gene was chosen for subsequent protein function research．The LvPT is 687 bp in length，encodes 228 amino acids，with a theoretical molecular weight of about 25．829 kDa． The fusion expression vector primers of the open reading frame CP,NS 1 and NS2 were designed according to a IHHNV Fujian strain(GenBank NC NC_002 1 90．2)， and yeast shuttle expression plasmid pGBKT7-CP, pGBKT7-NS 1，pGBKT7-NS2 and pGADT7一LvPT were constructed．The yeast shuttle plasmids that were detected by DDO／X／A and showed no self activation 万方数据  phenomenon could be used in protein interaction research．The recombinant strains Y2HGold(pGBKT7-C只pGBKT7-NS 1，pGBKT7-NS2) were respectively fused to the recombinant strains Y1 87(pGADT7-LvPT)，and all combinations were found to be positive by QDODUA filter．Further more， strong interactions were found by QDO／X／A filter between pGBKT7-CP and pGADT7-LvPT,pGBKT7-NS 1 and pGADT7-LvPT,res