Dye Charlotte, Siddell Stuart G
Department of Cellular and Molecular Medicine, Medical and Veterinary Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom.
J Feline Med Surg. 2007 Jun;9(3):202-13. doi: 10.1016/j.jfms.2006.12.002. Epub 2007 Mar 23.
This paper reports the first genomic RNA sequence of a field strain feline coronavirus (FCoV). Viral RNA was isolated at post mortem from the jejunum and liver of a cat with feline infectious peritonitis (FIP). A consensus sequence of the jejunum-derived genomic RNA (FCoV C1Je) was determined from overlapping cDNA fragments produced by reverse transcriptase polymerase chain reaction (RT-PCR) amplification. RT-PCR products were sequenced by a reiterative sequencing strategy and the genomic RNA termini were determined using a rapid amplification of cDNA ends PCR strategy. The FCoV C1Je genome was found to be 29,255 nucleotides in length, excluding the poly(A) tail. Comparison of the FCoV C1Je genomic RNA sequence with that of the laboratory strain FCoV FIP virus (FIPV) 79-1146 showed that both viruses have a similar genome organisation and predictions made for the open reading frames and cis-acting elements of the FIPV 79-1146 genome hold true for FCoV C1Je. In addition, the sequence of the 3'-proximal third of the liver derived genomic RNA (FCoV C1Li), which encompasses the structural and accessory protein genes of the virus, was also determined. Comparisons of the enteric (jejunum) and non-enteric (liver) derived viral RNA sequences revealed 100% nucleotide identity, a finding that questions the well accepted 'internal mutation theory' of FIPV pathogenicity.
本文报道了一株猫冠状病毒(FCoV)野毒株的首个基因组RNA序列。在尸检时从一只患有猫传染性腹膜炎(FIP)的猫的空肠和肝脏中分离出病毒RNA。通过逆转录聚合酶链反应(RT-PCR)扩增产生的重叠cDNA片段确定了源自空肠的基因组RNA(FCoV C1Je)的共有序列。通过迭代测序策略对RT-PCR产物进行测序,并使用cDNA末端快速扩增PCR策略确定基因组RNA末端。发现FCoV C1Je基因组长度为29,255个核苷酸,不包括聚(A)尾。将FCoV C1Je基因组RNA序列与实验室毒株FCoV FIP病毒(FIPV)79-1146的序列进行比较,结果表明这两种病毒具有相似的基因组结构,并且对FIPV 79-1146基因组的开放阅读框和顺式作用元件所做的预测对FCoV C1Je也适用。此外,还确定了源自肝脏的基因组RNA(FCoV C1Li)3'-近端三分之一的序列,该序列包含病毒的结构和辅助蛋白基因。对源自肠道(空肠)和非肠道(肝脏)的病毒RNA序列进行比较,发现核苷酸同一性为100%,这一发现对广为接受的FIPV致病性“内部突变理论”提出了质疑。