eb病毒相关疾病中eb病毒Bam HI - F DNA片段的序列分析

M Y Liu, K C Chen, I J Su, M M Hsu, J Y Chen, C S Yang
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引用次数: 0

摘要

为了比较EBV相关疾病中eb病毒(EBV) Bam HI - F DNA片段的核苷酸差异,采用聚合酶链反应(PCR)扩增了来自不同组织DNA的F片段的一个亚片段(核苷酸为55,381-56,020),包括20例npc、2例b细胞淋巴瘤、2例t细胞淋巴瘤、3例传染性单核细胞增多症(IM)和3例正常对照。采用PCR直接测序和DNA克隆后测序两种方法测定DNA序列。将PCR产物克隆到pGEM-3Z载体上,利用重组质粒转化DH5 α感受态细胞。从正确的转化子中制备质粒DNA进行DNA测序。结果表明,f变异在npc、b细胞淋巴瘤、t细胞淋巴瘤、IMs和正常人中的比例分别为40%、0%、0%、33%和33%。由于f变异体在鼻咽癌组织中并不比非肿瘤组织更普遍,我们推测f变异体与鼻咽癌之间没有很强的关联。这些结果与其他报告不同。在部分鼻咽癌患者和正常人中均发现F株和F变异体同时感染。对35株EBV分离株30个组织dna的Bam HI - F亚片段进行分析,发现B95-8株的4个相应位置发生了变化。它们是55,473位核苷酸T被G取代,55,543位核苷酸后插入TGT, 55,564位核苷酸A被G取代,55,958位核苷酸T被c取代。这4个核苷酸变化可能赋予了台湾菌株的特征。F株在55,519、55,596、55,680、55,703和55,895位点的核苷酸分别为T、T、A、A和C, F株的核苷酸分别为C、C、C、C和T,这两种模式与eb病毒A型和B型不相关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Sequence analyses of Epstein-Barr virus (EBV) Bam HI F DNA fragment in EBV associated diseases].

In order to compare nucleotide differences of Epstein-Barr virus (EBV) Bam HI F DNA fragment from various EBV associated diseases, polymerase chain reactions (PCR) were used to amplify a subfragment (nucleotides 55,381-56,020) of the F fragment from different tissue DNAs, including 20 NPCs, 2 B-cell lymphomas, 2 T-cell lymphomas, 3 infectious mononucleosis (IM), and 3 normal controls. DNA sequences were determined by PCR direct sequencing or sequencing after DNA cloning. The PCR products were cloned into pGEM-3Z vector, then the resulting recombinant plasmids were used to transform DH5 alpha competent cells. Plasmid DNAs from the correct transformants were prepared for DNA sequencing. The results showed that the proportion of the f variant in NPCs, B-cell lymphomas, T-cell lymphomas, IMs, and normals were 40%, 0%, 0%, 33%, and 33%, respectively. Because the f variant was not specifically more prevalent in NPC tissues compared to the non-tumor tissues, we speculate that there is no strong association between the f variant and NPC. These results were different from other reports. Coinfection of the F strain and the f variant was found both in some NPC patients and normal individuals. Analyses of Bam HI F subfragments of 35 EBV isolates from the 30 tissue DNAs revealed that there were changes at four corresponding positions of the B95-8 strain. They were nucleotide T at 55,473 replaced by G, an insertion of TGT after nucleotide 55,543, nucleotide A at 55,564 replaced by G, and nucleotide T at 55,958 replaced by C. These 4 nucleotide changes may confer a character of Taiwan strains. The nucleotides of the F strain at coordinates 55,519, 55,596, 55,680, 55,703, and 55,895 were T, T, A, A, and C, and those for the f variant were C, C, C, C, and T. These two patterns were not correlated with types A and B of EBV.

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