TYLCV-CHI侵染诱导的抗病毒GFP标记的载体构建

以CTAB法提取含有番茄黄化曲叶病毒TYLCV-CHI病原的烟草曲 叶病烟草DNA为模板，扩增该病毒外壳蛋白启动子序列，克隆到pGEM7Z载体上，在启动子下 游连接上能表达毒素蛋白的RNase基因和终止子NOS，最终将构建的基因转移到带有绿色荧光 蛋白GFP的植物表达载体上，构建了通过病毒侵染后激发病毒外壳蛋白启动子从而表达毒素 基因，产生毒素杀死病毒防止感染继续扩散的自我保护机制的基因工程植物表达载体。

There is the second virus that causes tobacco leaf curl disease (not TbLCV-CHI) in the field

Sequence analysis of virus isolation DNA of tobacco leaf curl disease shows that there is the second geminivirus (not Chinese Tobacco Leaf Curl Virus, TbLCV-CHI) that causes tobacco leaf curl disease in the field in the Guangxi Zhuang Autonomous Region, China. This virus DNA-A contains 2 734 nt. Large intergenic region (LIR) contains 269 nt, the virus sense strand contains 2 open reading frames (ORFs): AV1 (115 aa) and AV2 (coat protein gene, CP, 256 aa), and the complementary sense strand contains 4 ORFs: AC1 (replicase gene, 361 aa), AC2 (transactivator, 134 aa), ACS (134 aa) and AC4 (97 aa). The virus belongs to one kind of subgroup III gemini- viruses from old world, and could be the Chinese tomato yellow leaf curl virus (TYLCV-CHI).