Noiman S, Yaniv A, Sherman L, Tronick S R, Gazit A
Department of Human Microbiology, Sackler School of Medicine, Tel Aviv University, Israel.
J Virol. 1990 Apr;64(4):1839-43. doi: 10.1128/JVI.64.4.1839-1843.1990.
The pattern of expression of the equine infectious anemia virus (EIAV) genome in a persistently infected canine cell line was determined. Five EIAV-specific transcripts (8.2, 5.0, 4.0, 2, and 1.8 kilobases [kb]) were detected by using subgenomic restriction enzyme fragments of EIAV DNA and EIAV-specific oligonucleotides as probes. The 8.2-kb mRNA could be shown to represent viral genomic RNA, whereas the smaller transcripts were generated by splicing events. Evidence was obtained that indicated that each subgenomic RNA species shared a common 5'-splice donor. The 5.0-kb mRNA was found to be expressed at relatively low levels, was difficult to detect consistently, and appeared to be generated by a single splicing event which linked the 5' exon to the 3' region of pol. The 4.0-kb transcript was concluded to be the env mRNA on the basis of its hybridization pattern with the various probes and its abundance. The 2-kb species was found to be multiply spliced and was encoded by sequences derived from orf2 but was not detected by probes representing 3'-env/3'-orf sequences. The 1.8-kb species was shown to consist of sequences representing orf1, part of orf2, and the 3'-orf/env and may represent the message for the EIAV trans-activator gene.
确定了马传染性贫血病毒(EIAV)基因组在一个持续感染的犬类细胞系中的表达模式。通过使用EIAV DNA的亚基因组限制性酶切片段和EIAV特异性寡核苷酸作为探针,检测到了5种EIAV特异性转录本(8.2、5.0、4.0、2和1.8千碱基[kb])。8.2-kb的mRNA可被证明代表病毒基因组RNA,而较小的转录本是由剪接事件产生的。获得的证据表明,每个亚基因组RNA种类共享一个共同的5'-剪接供体。发现5.0-kb的mRNA表达水平相对较低,难以持续检测到,并且似乎是由一个单一的剪接事件产生的,该事件将5'外显子与pol的3'区域连接起来。基于其与各种探针的杂交模式及其丰度,推断4.0-kb的转录本为env mRNA。发现2-kb的种类是多重剪接的,由源自orf2的序列编码,但未被代表3'-env/3'-orf序列的探针检测到。1.8-kb的种类被证明由代表orf1、orf2的一部分以及3'-orf/env的序列组成,可能代表EIAV反式激活基因的信息。