Covey S N, Hull R
Department of Virus Research, John Innes Institute, Colney Lane, Norwich NR4 7UH, United Kingdom.
Virology. 1981 Jun;111(2):463-74. doi: 10.1016/0042-6822(81)90349-4.
We have detected several cauliflower mosaic virus transcripts in infected turnip leaves by northern-blot hybridization. These RNAs ranged in size from 0.9 to about 8 kb. Two species, a heterogeneous 7-to 8-kb RNA and a 2.3-kb RNA, accumulated radioactivity when CaMV-infected leaves were labeled with [32P]orthophosphate 20 days postinoculation. An abundant 62,000 MW polypeptide was synthesized in a rabbit reticulocyte lysate programmed with RNA from infected plants, but this polypeptide was absent when RNA from noninfected plants was used to direct translation. The 62,000 MW polypeptide was also the major in vitro product specified by virus-specific poly(A)+RNA purified by hybridization with CaMV DNA immobilized on DBM paper. The in vitro-synthesised 62,000 MW polypeptide was shown to be very similar to the major protein component of virus inclusion bodies by peptide fingerprint analysis. The 2.3-kb transcript is the mRNA encoding the 62,000 MW inclusion body protein. Crossed-contact hybridization mapping of this messenger on cloned CaMV DNA revealed that it is transcribed from the contiguous EcoR1 fragments d and b. The 7- to 8-kb RNA hybridized to all EcoR1 fragments and is probably a full-length primary transcript of the alpha-DNA strand.
通过Northern杂交,我们在受感染的芜菁叶片中检测到了几种花椰菜花叶病毒转录本。这些RNA的大小在0.9至约8 kb之间。接种20天后,用[32P]正磷酸盐标记CaMV感染的叶片时,两种RNA,即一种大小不均一的7至8 kb RNA和一种2.3 kb RNA积累了放射性。在用感染植物的RNA编程的兔网织红细胞裂解物中合成了一种丰富的62,000 MW多肽,但当使用未感染植物的RNA指导翻译时,这种多肽不存在。62,000 MW多肽也是通过与固定在DBM纸上的CaMV DNA杂交纯化的病毒特异性聚腺苷酸加尾RNA指定的主要体外产物。通过肽指纹分析表明,体外合成的62,000 MW多肽与病毒包涵体的主要蛋白质成分非常相似。2.3 kb转录本是编码62,000 MW包涵体蛋白的mRNA。该信使RNA在克隆的CaMV DNA上的交叉接触杂交图谱显示,它是从相邻的EcoR1片段d和b转录而来的。7至8 kb RNA与所有EcoR1片段杂交,可能是α-DNA链的全长初级转录本。