在豆科原生质体中检测双联病毒早期复制阶段的原位检测。

In situ detection of early replication phases of a gemini virus in legume protoplasts.

机构信息

Department of Genetics and Development, University of Illinois, 61801, Urbana, IL, USA.

出版信息

Plant Cell Rep. 1987 Dec;6(6):462-5. doi: 10.1007/BF00272783.

Abstract

Protoplasts of Phaseolus vulgaris L. (Top Crop), infected with bean golden mosaic virus, were isolated and fixed by various methods for in situ hybridization. An iodine-125 labeled probe was made from the replicative form of the virus. The localization and quantitation was done by autoradiography. Cell wall removal lowered the background and allowed a more accurate analysis. RNase was used to eliminate the possibility of hybrids to RNA. The evidence suggests a sequence of virus movements starting from rough endoplasm reticulum, moving to the nuclear membrane, and finally with the highest concentration inside the nucleus.

摘要

菜豆（顶级作物）原生质体感染菜豆花叶病毒后，采用不同方法固定并原位杂交。用病毒复制型制备放射性碘 125 标记探针。放射自显影进行定位和定量分析。去除细胞壁降低了背景，使分析更准确。用 RNase 消除了杂交到 RNA 的可能性。证据表明病毒的运动顺序是从粗糙内质网开始，移动到核膜，最终在核内浓度最高。