斜纹夜蛾spl14-3-3ζ基因的克隆 表达及其对莱氏野村菌侵染相关功能的分析-cloning and expression of spodoptera litura spl 14 - 3 - 3 ζ gene and analysis of its infection-related functions to nomura leguminicola.docxVIP

重庆大学 重庆大学硕士学位论文 英文摘要 PAGE PAGE VI ABSTRACT The 14-3-3 proteins, a highly evolutionarily conserved and ubiquitous protein family in eukaryotic cells, have a range of biological functions including regulation of signal transduction, stress response, apoptosis, and controlling of the cell cycle. To investigate the function of 14-3-3 in Spodoptera litura, the full length of 14-3-3ζ, named as SpL14-3-3ζ, was cloned from S. litura on the basis of an expressed sequence tag (EST) of 14-3-3ζ from the S. litura fat body suppression subtractive hybridization library (SSH). By qPCR and RNAi, we studied the function of SpL14-3-3ζ in the development of S. litura, and the molecular response relationship upon Nomuraea rileyi challenge. Our results will be helpful for understanding the molecular response mechanism of S. litura to N. rileyi infection, providing a scientific basis for controlling S. litura with the 14-3-3 protein as the target. ① Based on an EST of 14-3-3ζ from the S. litura SSH library, the full length of 14-3-3ζ was cloned from S. litura using SMART RACE and named SpL14-3-3ζ, whose amino acids sequence was identified by ORF Finder. The physical and chemical properties and the structure of the protein were predicted by Expasy etc. softwares. The homologous comparison of 14-3-3 proteins from various species was being done using DNAMAN. And then multiple alignment analysis and the phylogenetic tree were performed by ClustalW and MEGA software. The results showed that SpL14-3-3ζ cDNA was 1196bp long with an open reading frame 744bp encoding 247 amino acids. The molecular weight of the protein was 28.0kDa with isoelectric point of 4.6, without signal peptide or transmembrane domain. Multiple alignment analysis revealed the putative amino acids shared 80% homology with 14-3-3ζ from other organisms. Phylogenetic analysis confirmed SpL14-3-3ζ was closely related to other available lepidopteran 14-3-3ζ. Genome structure showed that SpL14-3-3ζ gene contained four exons, th