Sequence relatedness of Palyam virus genes to cognates of the Palyam serogroup viruses by RNA-RNA blot hybridization.
Sequence relatedness of Palyam virus genes to cognates of the Palyam serogroup viruses by RNA-RNA blot hybridization.
复制标题
通过 RNA-RNA 印迹杂交分析 Palyam 病毒基因与 Palyam 血清群病毒同源序列的相关性。
DOI：
10.1016/0042-6822(85)90096-0
复制
发表时间：
1985
期刊：
影响因子：
3.7
通讯作者：
Knudson,DL
中科院分区：
文献类型：
--
作者：
Bodkin,DK;Knudson,DL
环状病毒的Palyam血清组的9个成员的同源基因已经被鉴定，并且它们与原型Palyam病毒的相关性已经被确定。将病毒dsRNA片段通过10%聚丙烯酰胺凝胶电泳，转移到膜上，并在使用52°、50%甲酰胺、5X SSC的杂交条件下与来自Palyam病毒的标记RNA杂交。通过将其基因组与[5′-32 P] PCP标记的来自Palyam病毒的分离片段杂交，鉴定每个病毒分离株的同源基因。来自Palyam的单个片段与其他分离物中不超过一个片段杂交。10个基因中的9个在Palyam和其他8个分离株中的7个之间表现出核酸序列同源性。Palyam的基因2只与CSIRO Village的基因2杂交，且与血清型特异性相关。由于CSIRO Village是血清群中唯一在中和试验中与Palyam交叉反应的成员，因此基因2可能编码中和抗原。给定病毒中其余9个基因的杂交信号强度的变化表明，该组成员之间保守基因的数量和身份不同。基因5，7和9是最保守的基因的血清组的所有成员，而Palyam基因1，3，4，8，和10的相关性水平在其他分离株的同源物在这些杂交条件下变化。
Cognate genes of nine members of the Palyam serogroup of orbiviruses have been identified and their relatedness to the prototype, Palyam virus, has been determined. Viral dsRNA segments were electrophoresed through 10% polyacrylamide gels, transferred to membranes, and hybridized to labeled RNA from Palyam virus under hybridization conditions using 52°, 50% formamide, 5X SSC. Cognate genes of each virus isolate were identified by hybridizing their genomes to [5′-32P]pCp-labeled, isolated segments from Palyam virus. Single segments from Palyam hybridized to no more than one segment in the other isolates. Nine of the 10 genes exhibited nucleic acid sequence homology between Palyam and seven of the other eight isolates. Gene 2 of Palyam hybridized only with gene 2 of CSIRO Village, and it was correlated with serotype specificity. Since CSIRO Village is the only member of the serogroup which cross-reacts with Palyam in neutralization tests, gene 2 may encode the neutralization antigen. Variation in the intensity of the hybridization signals of the remaining nine genes within a given virus indicated that the number and identity of conserved genes differed between members of the group. Genes 5, 7, and 9 were the most conserved genes for all members of the serogroup, while the levels of relatedness of Palyam genes 1, 3, 4, 8, and 10 to their cognates in the other isolates varied under these hybridization conditions.