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在一组相关的爱泼斯坦-巴尔病毒家族成员中,基因组质粒大小存在显著差异。

Marked variation in the size of genomic plasmids among members of a family of related Epstein-Barr viruses.

作者信息

Kolman J L, Kolman C J, Miller G

机构信息

Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06510.

出版信息

Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7772-6. doi: 10.1073/pnas.89.16.7772.

Abstract

Epstein-Barr virus (EBV) genomes in the P3J-HR-1 (HR-1) Burkitt lymphoma cell line rearrange at a high rate. Previously described deletions and rearrangements in HR-1 cells have been found at sites of EBV replication in vivo, suggesting that DNA rearrangement may be an integral aspect of EBV biology and pathogenesis. We examined the structure of linear EBV genomes in subcultures of HR-1 cells using contour-clamped homogenous electric field gel electrophoresis. We developed a second pulsed electrophoretic technique to separate intracellular circular EBV plasmids. The standard, linear HR-1 EBV genome was approximately 155 kilobases in length. Linear molecules of less than unit length, presumably defective genomes, were seen in numerous subcultures. Linear intracellular genomes greater than 155 kilobases were also detected, but only linear genomes of 155 kilobases or less were packaged into virions. The size of circular EBV plasmids also varied greatly among HR-1 subcultures, some of which contained two plasmids of different size. The progeny of the unusual circular plasmids could be either standard or nonstandard linear genomes. No aberrant linear or circular form was detected in a subculture carrying the previously described het fragments. Pulsed-gel electrophoresis has provided two additional characteristics of mutant EBVs: abnormal linear and circular genome configurations.

摘要

P3J-HR-1(HR-1)伯基特淋巴瘤细胞系中的爱泼斯坦-巴尔病毒(EBV)基因组以高频率发生重排。此前在HR-1细胞中描述的缺失和重排已在EBV体内复制位点被发现,这表明DNA重排可能是EBV生物学和发病机制的一个重要方面。我们使用轮廓夹均等电场凝胶电泳检测了HR-1细胞亚培养物中线性EBV基因组的结构。我们开发了第二种脉冲电泳技术来分离细胞内的环状EBV质粒。标准的线性HR-1 EBV基因组长度约为155千碱基对。在许多亚培养物中都观察到了长度小于单位长度的线性分子,推测为缺陷基因组。还检测到了长度大于155千碱基对的线性细胞内基因组,但只有155千碱基对或更短的线性基因组被包装进病毒粒子。HR-1亚培养物中环状EBV质粒的大小也有很大差异,其中一些含有两种不同大小的质粒。这些异常环状质粒的后代可能是标准的或非标准的线性基因组。在携带先前描述的het片段的亚培养物中未检测到异常的线性或环状形式。脉冲凝胶电泳揭示了突变EBV的另外两个特征:异常的线性和环状基因组构型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae15/49793/7772ce109070/pnas01090-0495-a.jpg

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