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由感染黄瓜花叶病毒(CMV)、健康和转外壳蛋白(CP)基因的烟叶以原生质体法制备纯化的叶绿体。经SDS—PAGE电泳,银染色,比较其蛋白质图谱。用CMV外壳蛋白游离亚基制备的抗血清进行Western blotting。结果发现:1.CP存在于CMV侵染的烟叶绿体中。2.叶绿体中CP的浓度和花叶症状严重程度呈正相关;3.表达CP的转基因烟草叶绿体中未测出CP存在。根据以上结果,认为CMV侵染的烟花叶症状产生与CP进入叶绿体有直接相关性。
Purified chloroplasts were prepared by the protoplast method from tobacco leaf infected with cucumber mosaic virus (CMV), health and coat protein (CP) genes. After SDS-PAGE electrophoresis, silver staining, compared with the protein map. Western blotting was performed using antisera prepared from the CMV coat protein free subunit. The results showed that: 1.CP exists in CMV infected tobacco chloroplasts. The concentration of CP in chloroplast was positively correlated with the severity of mosaic symptom.3. No CP was detected in the chloroplast of transgenic tobacco expressing CP. Based on the above results, it is considered that the CMV infected tobacco leaf symptoms are directly related to the CP entry into the chloroplast.