凋亡抑制蛋白基因dsRNA转基因烟草表达载体的构建

Construction of Transgenic Tobacco Plant Vector for Inhibitor of Apoptosis Protein Gene dsRNA

为了利用RNA干扰技术防治烟粉虱,需要构建凋亡抑制蛋白(IAP)基因dsRNA转基因烟草表达载体。利用PCR技术扩增烟粉虱IAP基因,将其连接到pMD18-T载体中,用BglⅡ、XhoⅠ和BamHⅠ、SalⅠ分别酶切,将获得的片段分别反向、正向连接至pUC-RNAi载体,用PstⅠ酶切pUC-RNAi-IAP,回收酶切片段,将其连接到表达载体pCAMBIA-2300-35S中,并进行酶切验证。IAPdsRNA转基因烟草表达载体的成功构建为下一步转基因烟草防治烟粉虱的研究奠定基础。

To contol Bemisia tabaci by RNAi, transgenic tobacco plant vector for inhibitor of apoptosis protein （lAP） gene dsRNA should be constructed. In this study, we amplified the IAP gene of Bemisia tabaci by PCR, inserted it into pMD18-T, digested with BglII/XhoI or BamHI/SalI respectively, conducted the acquired fragments to pUC-RNAi vector, digested with PstI, and cloned into the vector pCAMBIA-2300-35S. Transgenic tobacco plant vector was confirmed by PstI digesting. The construction of transgenic to- bacco plant vector for IA P dsRNA would lay the foundation for the following study of pest control.