摘要
我们曾报道,十字花科萝卜的各种栽培变种都含有抗病毒、抗癌作用的干扰素诱生剂,其有效成分为dsRNA。本实验构建了含有心里美萝卜dsRNA的cDNA重组质粒。以其cDNA为探针进行杂交试验,探讨各种变种萝卜dsRNA之间的亲缘关系。 萝卜dsRNA的分离和纯化参照文献,先从萝卜提取总核酸,然后经CF11纤维素柱层析和低融点琼脂糖制备电泳,获得较纯的dsRNA。
The dsRNA isolated from the root of Raphanus sativus ( cv. Xin li mei)was reverse-trancribed into cDNA and cloned into pBR322.The pBR 322 clone with largest cDNA insert was chosen for the preparation of probe for dot blot bybridization assay. The cDNA was labeled by randomly primed incorporation of digoxigenin-labeled-dUTP. After hybridization to the target nucleic acid,the hybrids were detected by enzyme linked immunoassay using antibodyconjugate.The labeled cDNA probe hybridized strongly with the dsRNA isolated from the root of the different cultvars of P.sativus. These results suggest that there is sequence homology between dsRNA from different cultivars. However, it remains to be clarified whether or not the dsRNA is a transcriptional product of chromosome DNA of the plant.
出处
《病毒学报》
CAS
CSCD
北大核心
1992年第4期364-366,共3页
Chinese Journal of Virology
关键词
DSRNA
萝卜
CDNA
杂交分析
克隆
Cloning of dsRNA Raphanus sativus-root cDNA hybridization assay
