Mayo M A
Intervirology. 1982;17(4):240-6. doi: 10.1159/000149294.
Tobacco protoplasts were irradiated with UV light, inoculated with tobacco rattle virus (TRV), and then cultured in media containing 35S-methionine. Polypeptides induced by TRV infection were detected by PAGE of protein extracts. Three 35S-labeled polypeptides (apparent molecular weights 187,000, 142,000, and 31,000) occurred in extracts of virus-infected protoplasts but not in extracts of buffer-inoculated protoplasts. The 31,000 mol. wt. polypeptides co-migrated with TRV coat protein in polyacrylamide gels and was not detected when inocula contained only the longer particle of TRV. The 187,000 and 142,000 mol. wt. polypeptides migrated in positions very close to products of in vitro translation of TRV RNA-1. Experiments in which 35S-methionine was supplied at different times during multiplication suggested that the bulk of translation of RNA-1 occurred sooner after inoculation than did that of RNA-2.
用紫外线照射烟草原生质体，接种烟草脆裂病毒（TRV），然后在含有35S-甲硫氨酸的培养基中培养。通过对蛋白质提取物进行聚丙烯酰胺凝胶电泳（PAGE）来检测由TRV感染诱导的多肽。在病毒感染的原生质体提取物中出现了三种35S标记的多肽（表观分子量分别为187,000、142,000和31,000），而在缓冲液接种的原生质体提取物中未出现。分子量为31,000的多肽在聚丙烯酰胺凝胶中与TRV外壳蛋白迁移位置相同，当接种物仅包含TRV的较长颗粒时未检测到该多肽。分子量为187,000和142,000的多肽迁移位置与TRV RNA-1体外翻译产物非常接近。在繁殖过程中不同时间供应35S-甲硫氨酸的实验表明，RNA-1的大部分翻译在接种后比RNA-2发生得更早。
