杆状病毒基因转导载体具有安全、高效和容量大等优点，但介导杆状病毒基因组加工和压缩包装的顺式作用元件仍未被发现，影响其转导载体的改良和应用。申请人在近期首次发现了为杆状病毒核衣壳装配所必需的顺式作用元件NAE。前期研究证明，NAE不参与病毒基因组的合成。因此，我们提出一个合理的推测：NAE可能通过参与杆状病毒基因组在合成后的加工或压缩包装过程，从而介导核衣壳的装配。为此，本课题将以苜蓿丫纹夜蛾核多角体病毒为材料，构建NAE缺失型重组病毒，通过脉冲场电泳、qPCR、电子显微镜观察和iTRAQ蛋白质谱等方法，针对性地检测NAE对病毒基因组的构型和包装效率的影响，以鉴定NAE在病毒基因组线性化、环化、同源重组和压缩包装过程中的功能。研究结果对阐明杆状病毒及其他大型环状DNA病毒核衣壳装配机制有重要意义。同时，本课题的研究成果也可为改良杆状病毒基因转导载体提供理论基础。

The baculovirus gene transduction vector has many special advantages on safety, transduction efficacy, and foreign gene capacity. However, the cis-acting element that mediates the baculovirus genome processing and encapsidation was not yet discovered, which hinders the improvement and application of the baculovirus vector. We recently identified for the first time a cis-acting element NAE in the baculovirus genome, which is essential for the viral nucleocapsid assembly. It has been demonstrated that NAE is not required for the viral genome synthesis. Therefore, it is a reasonable hypothesis that NAE mediate the baculovirus nucleocapsid assembly via its participation in the genome processing or encapsidation after viral genome synthesis. To this end, with the Autographa californica multiple nucleopolyhedrovirus as material, this project is going to detect the influence of NAE on the viral genome configuration and the encapsidation efficiency, in order to determine the function of NAE in the linearization, circularization, homologous recombination and encapsidation of the baculovirus genome. Methods including construction of NAE-deletion virus, pulse-field electrophoresis, real-time PCR, electron microscopy, and iTRAQ mass spectrometry will be utilized. The results will be important for the research on the nucleocapsid assembly mechanism of baculovirus as well as other circular DNA viruses, and it will also provide an important reference for the improvement of baculovirus gene transduction vector.