THE ROLE OF REPETITIVE DNA-SEQUENCES IN THE SIZE VARIATION OF EPSTEIN-BARR-VIRUS (EBV) NUCLEAR ANTIGENS, AND THE IDENTIFICATION OF DIFFERENT EBV ISOLATES USING RFLP AND PCR ANALYSIS
THE ROLE OF REPETITIVE DNA-SEQUENCES IN THE SIZE VARIATION OF EPSTEIN-BARR-VIRUS (EBV) NUCLEAR ANTIGENS, AND THE IDENTIFICATION OF DIFFERENT EBV ISOLATES USING RFLP AND PCR ANALYSIS
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DOI：
10.1099/0022-1317-76-4-779
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发表时间：
1995-04-01
影响因子：
3.8
通讯作者：
ERNBERG, I
中科院分区：
文献类型：
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作者：
FALK, K;GRATAMA, JW;ERNBERG, I
6种EB病毒（EBV）核抗原蛋白（EBNA-1-6）在不同病毒分离株之间显示出特征性的大小差异;这是流行病学研究（Ebnotyping）中用于鉴定病毒分离株来源的特征。我们现在已经研究了编码EBNA的外显子内的限制性片段长度多态性（RFLP）与相应蛋白质的分子量之间的相关性。B 95 -8 EBV株用作原型病毒。EBNA-1、EBNA-3和EBNA-6的表观分子量变化与编码这些多肽中重复序列的RFLP大小呈正相关。EBNA-2的表观分子量和重复序列长度之间没有相关性，EBNA-4的表观分子量和RFLP大小在重复序列上几乎没有变化。基于EBNA-6的表观分子量和RFLP大小之间的强相关性，我们开发了区分不同EBV分离株的EBNA-6 PCR检测。该测定提供了使用未培养的材料（例如咽喉洗涤液、血液或活检）进行EBV表征的优点，从而避免了在建立蛋白质水平的Ebnotyping所需的淋巴母细胞系期间发生的针对转化不良的菌株的选择。
The six Epstein-Barr virus (EBV) nuclear antigen proteins (EBNA-1-6) show characteristic size variations between different virus isolates; this is a feature that has been used to identify the source of virus isolates in epidemiological studies (Ebnotyping). We have now studied the correlation between restriction fragment length polymorphisms (RFLPs) within exons coding for the EBNAs and the molecular masses of the respective proteins. The B95-8 EBV strain was used as the prototype virus. The variation in apparent molecular mass of EBNA-1, -3 and -6 correlated positively with the size of RFLP coding for repeat sequences in these polypeptides. For EBNA-2, no correlation between apparent molecular mass and length of the repetitive sequences was found. The EBNA-4 protein showed virtually no variation in apparent molecular mass and RFLP size across the repeat sequence. Based on the strong correlation between apparent molecular mass and RFLP size for EBNA-6, we developed an EBNA-6 PCR assay that discriminated between different isolates of EBV. This assay offers the advantage of EBV characterization using uncultured material (e.g. throat washings, blood or biopsies), thus avoiding the selection against poorly transforming strains that occurs during establishment of lymphoblastoid cell lines required for Ebnotyping at the protein level.