Hilf M E, Karasev A V, Pappu H R, Gumpf D J, Niblett C L, Garnsey S M
USDA-ARS Horticultural Research Laboratory, Orlando, Florida 32803, USA.
Virology. 1995 Apr 20;208(2):576-82. doi: 10.1006/viro.1995.1188.
Citrus tristeza virus (CTV) specific RNAs extracted from infected citrus tissue were analyzed by Northern blot hybridization. RNAs were characterized by size and identified using cDNA probes specific to nine open reading frames (ORFs) identified by the analysis of sequence obtained from cDNA clones of the T36 isolate of CTV. Sequence specific cDNA probes identified the genomic RNA as well as subgenomic RNAs representing the p33, p65, p61, p27, p25, p18, p13, p20, and p23 ORFs in extracts of total or double-stranded RNA (dsRNA) isolated from infected tissue. A probe derived from the 3' terminal ORF (p23) hybridized to each of these subgenomic RNAs, indicating that the RNAs are 3' coterminal. The relative amounts of the different subgenomic RNAs varied widely. The RNAs for the p20 and p23 ORFs were the most abundant and surpassed the amount of the p25 or capsid protein specific subgenomic RNA. The number and sizes of the CTV subgenomic RNAs were the same in total RNA and dsRNA preparations. Propagation of T36 in seven different citrus hosts did not alter the pattern of subgenomic RNAs.
通过Northern印迹杂交分析从感染柑橘组织中提取的柑橘衰退病毒（CTV）特异性RNA。RNA通过大小进行表征，并使用针对CTV T36分离株cDNA克隆序列分析鉴定的9个开放阅读框（ORF）的特异性cDNA探针进行鉴定。序列特异性cDNA探针鉴定了基因组RNA以及代表从感染组织中分离的总RNA或双链RNA（dsRNA）提取物中p33、p65、p61、p27、p25、p18、p13、p20和p23 ORF的亚基因组RNA。源自3'末端ORF（p23）的探针与这些亚基因组RNA中的每一个杂交，表明这些RNA是3'共末端的。不同亚基因组RNA的相对含量差异很大。p20和p23 ORF的RNA最为丰富，超过了p25或衣壳蛋白特异性亚基因组RNA的量。CTV亚基因组RNA在总RNA和dsRNA制剂中的数量和大小相同。T36在七种不同柑橘宿主中的繁殖并未改变亚基因组RNA的模式。
