论文部分内容阅读
利用PCR方法克隆了香蕉束顶病毒中国漳州分离物(BBTV-ZZ)DNA 4编码区,序列分析结果表明该编码区由351个核苷酸组成,推测其编码是一个含116个氨基酸的蛋白质,利用植物双元载体pBin438构建了含有BBTV-ZZ DNA 4编码区的植物组成型表达载体pBBTV-4B,经农杆菌介导转化了三生烟(Nicotiana tabacum Xanthi nc),在防虫条件下,将运动缺陷型CMV-Fny突变株(CMV-Fny-△MP)人工接种到转基因植株下部叶片上,12d后,在转基因植株的非接种叶上显现出不同程度的系统侵染症状,间接异种动物双抗体夹心酶联免疫吸附方法(DAS-ELISA)检测结果显示在转基因植株的上部非接种叶中有CMV-Fny的积累,而非转基因对照植株组的上部非接种叶中始终没有显现系统侵染症状,DAS-ELISA检测也未见CMV-Fny的积累,这些实验结果表明转基因植株能够支持CMV-Fny-△Mp从细胞到细胞和长距离运动并形成系统侵染症状,由此推测BBTV-ZZ DNA 4编码的蛋白质可能具有运动蛋白功能。
The DNA 4 coding region of banana bunchy top virus China Zhangzhou isolate (BBTV-ZZ) was cloned by PCR. The sequence analysis showed that the coding region consisted of 351 nucleotides. The deduced amino acid sequence was encoded by a 116 amino acid protein, The plant constitutive expression vector pBBTV-4B containing the BBTV-ZZ DNA 4 coding region was constructed by using the plant binary vector pBin438, and Nicotiana tabacum Xanthi nc was transformed by Agrobacterium tumefaciens. Under the insect-resistant conditions, the movement The transgenic CMV-Fny mutant (CMV-Fny-△ MP) was inoculated on the lower leaves of transgenic plants. After 12 days, CMV-Fny mutant showed different degrees of systemic infection on non-inoculated leaves of transgenic plants. Indirect heterologous diabody The results of sandwich enzyme-linked immunosorbent assay (DAS-ELISA) showed that there was accumulation of CMV-Fny in the upper non-inoculated leaves of the transgenic plants, while the systemic non-inoculated leaves in the upper non-inoculated leaves of the non- No accumulation of CMV-Fny was detected by DAS-ELISA. These experimental results indicated that the transgenic plants could support CMV-Fny-△ Mp from cells to cells and long-distance movement and form systemic infection symptoms, suggesting that BBTV The protein encoded by ZZ DNA 4 may have motor protein function.