STRUCTURE AND FUNCTION OF THE ALEUTIAN MINK DISEASE PARVOVIRUS GENOME

这个项目的持续目的是确定基因组差异 在阿留申水貂病细小病毒（ADV）的各种毒株中， 将这些差异与病毒宿主范围和致病性联系起来。 研究了来自犹他州正在发生的ADV感染暴发的病毒。 采用PCR方法分离并鉴定161 bp的核心高变区 衣壳蛋白基因的区域。 多个样本的序列 水貂和野生浣熊在这次爆发中的关系非常密切 彼此相关（>97%）。 此外，他们更接近 与1型非致病性ADV-G相关（>97%），而与致病性 菌株如2型ADV-Utah、3型Danish ADV-K或 分离自野生臭鼬（均<90%）。病毒的扩展测序 来自这次爆发（ADV-TR）表明，在其他残留物， 衣壳蛋白基因与ADV-G不同， 因此，具有1型高变的病毒 区域可能是致病性的，然而，这些结果表明， 致病性和宿主范围的决定因素可能极其复杂。 ADV-TR对阿留申基因型和非阿留申基因型均具有高致病性 貂皮。ADV的致病株，包括ADV-TR，诱导抗病毒 抗体和病毒血症，但没有证据表明浣熊的病理。 ADV-TR 可能会从受感染的浣熊传给貂因此，浣熊可能会玩 在ADV的维持和传播中起作用。 其他旨在检查转录单位功能的工作， 衣壳蛋白P36启动子的研究。 第一链cDNA 已经从用几种细小病毒转染的细胞制备 用于5 '-RACE以比较转录的启动子构建体 在组成型和反式激活条件下的起始位点。 的 5 '-RACE片段已被克隆并正在表征。

The ongoing purpose of this project is to identify genomic differences among various strains of Aleutian mink disease parvovirus (ADV) and to relate these differences to viral host range and pathogenicity. Viruses from an ongoing outbreak of ADV infection in Utah were studied. PCR was used to isolate and characterize the 161 bp core hypervariable region of the capsid protein gene. Sequence of specimens from multiple mink and feral raccoons involved in the outbreak were very closely related to each other (>97%). Furthermore, they were more closely related to the type 1 nonpathogenic ADV-G (>97%) than to pathogenic strains such as the type 2 ADV-Utah, the type 3 Danish ADV-K or an isolate from feral skunks (all <90%). Expanded sequencing of a virus derived from this outbreak (ADV-TR) indicated that at other residues, the capsid protein gene differed from ADV-G and more closely resembled pathogenic strains of ADV. Thus, a virus with a type 1 hypervariable region can be pathogenic, however, these results indicate that viral determinants for pathogenicity and host range may be extremely complex. ADV-TR was highly pathogenic for both Aleutian and non-Aleutian genotype mink. Pathogenic strains of ADV, including ADV-TR, induced antiviral antibodies and viremia, but no evidence of pathology in raccoons. ADV-TR could be passaged from infected raccoons to mink. Thus, raccoons may play a role in maintenance and transmission of ADV. Other work designed to examine function of the transcription unit from the capsid protein P36 promoter has been undertaken. First strand cDNAs have been prepared from cells transfected with several parvovirus promoter constructs for use in 5'-RACE to compare the transcription initiation site under constitutive and trans-activated conditions. The 5'-RACE fragments have been cloned and are being characterized.