GENOME ORGANIZATION OF A PARTITIVIRUS FROM THE FILAMENTOUS ASCOMYCETE ATKINSONELLA HYPOXYLON
GENOME ORGANIZATION OF A PARTITIVIRUS FROM THE FILAMENTOUS ASCOMYCETE ATKINSONELLA HYPOXYLON
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DOI：
10.1099/0022-1317-76-6-1461
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发表时间：
1995-06-01
影响因子：
3.8
通讯作者：
HILLMAN, BI
中科院分区：
文献类型：
--
作者：
OH, CS;HILLMAN, BI
我们已经确定了丝状子囊菌Atkinsonella hypoxylon的几个分离株中的病毒。选择来自真菌2H的一个分离株的病毒进行基因组表征。纯化的病毒颗粒含有三种dsRNA，其大小通过凝胶电泳估计为2.2、2.1和1.8kb。合成、绘制并测序了代表分离株2 H的三个双链RNA片段的cDNA克隆库。通过北方印迹分析确定这三个片段彼此没有显著的相似性，并且通过核苷酸序列分析确定其大小为2180、2135和1790 nt。从dsRNA 1（分子量78 kDa）和2（74 kDa）的序列推导出长开放阅读框，但不从dsRNA 3推导。dsRNA 1和dsRNA 2的两个末端区域具有相似的核苷酸序列，如从5'RACE克隆所确定的。从dsRNA 1推导的氨基酸序列的比较揭示了与病毒RNA依赖性RNA聚合酶的相似性。通过聚丙烯酰胺凝胶分析，代表dsRNA 1和2的全长cDNA克隆的体外翻译各自产生> 70 kDa的单一主要产物。根据其双链RNA片段的特性，A. hypoxylon菌株2H属于偏粒病毒科，并且代表了该家族的第一个在一级核苷酸序列水平上完全表征的来自真菌宿主的成员。
We have identified viruses in several isolates of the filamentous ascomycete Atkinsonella hypoxylon. The virus from one isolate of the fungus, 2H, was selected for genomic characterization. Purified virus particles contained three dsRNAs with sizes estimated by gel electrophoresis to be 2.2, 2.1 and 1.8 kb. A library of cDNA clones representing the three dsRNA segments of isolate 2H was synthesized, mapped and sequenced. The three segments had no significant similarity to each other, as determined by Northern blot analysis, and had sizes of 2180, 2135 and 1790 nt as determined by nucleotide sequence analysis. Long open reading frames were deduced from the sequences of dsRNAs 1 (molecular mass 78 kDa) and 2 (74 kDa), but not from dsRNA 3. Both terminal regions of dsRNA 1 and dsRNA 2 had similar nucleotide sequences, as determined from 5' RACE clones. Comparisons of the amino acid sequence deduced from dsRNA 1 revealed similarities with viral RNA-dependent RNA polymerases. Translation in vitro of full-length cDNA clones representing dsRNAs 1 and 2 each yielded single major products of > 70 kDa by analysis on polyacrylamide gels. Based on properties of its dsRNA segments, the virus of A. hypoxylon strain 2H fits into the Partitiviridae family, and represents the first member of this family from a fungal host completely characterized at the level of primary nucleotide sequence.