Molecular characterization of Drosophila C virus isolates
Molecular characterization of Drosophila C virus isolates
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DOI：
10.1006/jipa.1998.4830
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发表时间：
1999-05-01
影响因子：
3.4
通讯作者：
Christian, PD
中科院分区：
文献类型：
--
作者：
Johnson, KN;Christian, PD
采用逆转录聚合酶链式反应和限制性内切酶分析相结合的方法，对12株不同地区的果蝇C病毒分离株进行了鉴定。从cDNA中扩增出一条1.2kb的片段，用20种限制性内切酶进行酶切，得到图谱。对限制性片段数据的分析给出了分离株之间核苷酸差异的估计为0-10%。根据限制数据得出的遗传距离估计对这些分离物进行了分组。对于可以提纯单一基因的分离株，确定了病毒基因类型分布的地理模式。这4个摩洛哥分离株之间的亲缘关系非常近，只有一个限制性内切酶图谱不同。这两个澳大利亚分离株是彼此最近的亲缘关系，法国首次发现的两个分离株也是如此。本研究中所使用的PCR-RFLP技术为我们提供了一种简单的方法，可用于DCV分离株的特征分析。一个单一的酶，TAG I，产生了5种不同的和诊断的限制性片段模式，这使得分离株可以很容易地分配到本研究中确定的五种病毒基因型之一。(C)1999年学术出版社。
Reverse transcription coupled with polymerase chain reaction and restriction enzyme analysis was used to characterize 12 Drosophila C virus isolates from geographically different regions. A 1.2-kb fragment was amplified from cDNA and profiles from digestion with 20 restriction enzymes were generated. Analysis of the restriction fragment data gave estimates of nucleotide divergence of 0-10% between isolates. The isolates were grouped on the basis of genetic distance estimates derived from the restriction data. For the isolates from which a single genotype could be purified, a geographical pattern in the distribution of viral genotypes was identified. The 4 Moroccan isolates were very closely related to each other, differing in only 1 restriction profile. The 2 Australian isolates were each other's closest relatives, as were the 2 isolates first recovered in France. The PCR-RFLP technique used in this study has provided us with a simple procedure which can be used to characterize DCV isolates. A single enzyme, Tag I, generated 5 distinct and diagnostic restriction fragment patterns, which allowed easy assignment of isolates to one of the five viral genotypes identified in this study. (C) 1999 Academic Press.