Zuidema D, Heaton L A, Jackson A O
Department of Plant Pathology, University of California, Berkeley, California 94720, USA.
Virology. 1987 Aug;159(2):373-80. doi: 10.1016/0042-6822(87)90476-4.
The structure of the gene adjacent to the "leader RNA" gene of sonchus yellow net virus (SYNV), a plant rhabdovirus, was deduced by dideoxyribonucleotide sequence analysis of SYNV genomic (g) RNA and a series of plasmids constructed from SYNV gRNA or polyadenylated [poly(A)+] RNA from SYNV-infected plants. Evidence that this gene encodes the nucleocapsid (N) protein was obtained by reaction of SYNV N protein with polyclonal antibodies raised against recombinant proteins derived from the cloned gene. Experiments in which defined oligodeoxyribonucleotides were used to initiate reverse transcription of poly(A)+ RNA from SYNV-infected tobacco revealed that the N protein messenger (m) RNA gene begins at position 147 from the 3' end of the SYNV genome. Inspection of the sequence shows that this mRNA has a 56 nucleotide (NT) untranslated region followed by a 1425 NT open reading frame that is terminated by tandem UAA stop codons at positions 1628 to 1633 relative to the 3' end of SYNV gRNA. Little direct sequence homology is evident between the 475 amino acid polypeptide predicted from the SYNV sequence and the nucleocapsid (N) proteins deduced from nucleotide sequences of the Indiana and New Jersey serotypes of vesicular stomatitis virus (VSV) and rabies virus. However, a short region of possible importance contains a small group of chemically similar amino acids common to all four N proteins.
通过对南芥菜花叶病毒(SYNV,一种植物弹状病毒)基因组(g)RNA以及一系列由SYNV gRNA或感染SYNV的植物中的聚腺苷酸化[poly(A)+] RNA构建的质粒进行双脱氧核糖核苷酸序列分析,推导了与SYNV“前导RNA”基因相邻的基因结构。通过SYNV核衣壳(N)蛋白与针对克隆基因衍生的重组蛋白产生的多克隆抗体的反应,获得了该基因编码核衣壳(N)蛋白的证据。使用特定的寡脱氧核苷酸起始来自感染SYNV的烟草的poly(A)+ RNA的逆转录实验表明,N蛋白信使(m)RNA基因从SYNV基因组3'端的第147位开始。对序列的检查表明,该mRNA具有一个56个核苷酸(NT)的非翻译区,其后是一个1425 NT的开放阅读框,该开放阅读框在相对于SYNV gRNA 3'端的第1628至1633位由串联的UAA终止密码子终止。从SYNV序列预测的475个氨基酸的多肽与从水疱性口炎病毒(VSV)和狂犬病病毒的印第安纳和新泽西血清型的核苷酸序列推导的核衣壳(N)蛋白之间几乎没有明显的直接序列同源性。然而,一个可能重要的短区域包含所有四种N蛋白共有的一小群化学性质相似的氨基酸。