Stettler M, Zurbriggen A
Institute of Animal Neurology, University of Berne, Switzerland.
Vet Microbiol. 1995 May;44(2-4):211-7. doi: 10.1016/0378-1135(95)00014-2.
Virus persistence is essential in the chronic inflammatory canine distemper virus (CDV)-induced demyelinating disease. In the case of CDV there is a close association between persistence and virulence. Virulent CDV isolated from dogs with distemper shows immediate persistence in primary dog brain cell cultures (DBCC) and in different cell lines. We have evidence that the nucleocapsid (NP) protein plays an important role in the development of persistence. The NP-protein, the most abundant structural virus protein, also influences virus assembly and has some regulatory functions in virus transcription and replication. In this study we compared the nucleotide and deduced amino acid sequence of a virulent CDV strain (A75/17-CDV) to a culture-attenuated non-virulent strain (OP-CDV). Viral RNA was extracted from DBCC infected with virulent CDV. Virulent CDV retains its in vivo properties, such as virulence and ability to cause demyelination, when propagated in these DBCC. The viral RNA was reverse transcribed and the resulting cDNA amplified by polymerase chain reaction for subsequent cloning. The nucleotide sequences of these clones were determined by the dideoxy chain termination method. The number of nucleotides and the putative NP-protein of the virulent strain matched the attenuated CDV strain. We observed a total of 105 nucleotide differences. Three were localised within the 3' and five within the 5' non-coding region of the NP-gene. The 97 nucleotide changes within the coding region resulted in 22 amino acid differences. 10 of these amino acid (AA) modifications were within the N-terminal region (AA 1 to 159) and 12 within the C-terminal area (AA 351 to 523).(ABSTRACT TRUNCATED AT 250 WORDS)
病毒持续性在慢性炎症性犬瘟热病毒(CDV)诱导的脱髓鞘疾病中至关重要。就CDV而言,持续性与毒力密切相关。从患有犬瘟热的狗身上分离出的强毒株CDV在原代犬脑细胞培养物(DBCC)和不同细胞系中表现出立即持续性。我们有证据表明核衣壳(NP)蛋白在持续性的发展中起重要作用。NP蛋白是病毒中含量最丰富的结构蛋白,也影响病毒组装,并在病毒转录和复制中具有一些调节功能。在本研究中,我们将强毒株CDV(A75/17 - CDV)的核苷酸和推导的氨基酸序列与一株经培养减毒的无毒株(OP - CDV)进行了比较。从感染强毒株CDV的DBCC中提取病毒RNA。当在这些DBCC中繁殖时,强毒株CDV保留其体内特性,如毒力和引起脱髓鞘的能力。将病毒RNA反转录,所得cDNA通过聚合酶链反应扩增以用于后续克隆。通过双脱氧链终止法确定这些克隆的核苷酸序列。强毒株的核苷酸数量和推定的NP蛋白与减毒CDV株匹配。我们总共观察到105个核苷酸差异。其中三个位于NP基因的3'端非编码区内,五个位于5'端非编码区内。编码区内的97个核苷酸变化导致22个氨基酸差异。这些氨基酸(AA)修饰中有10个位于N端区域(第1至159位氨基酸),12个位于C端区域(第351至523位氨基酸)。(摘要截断于250字)