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爱泼斯坦-巴尔病毒经常整合到伯基特淋巴瘤细胞系(拉吉细胞系)的4号、2号、1号和7号染色体长臂上。

Epstein-Barr virus integrates frequently into chromosome 4q, 2q, 1q and 7q of Burkitt's lymphoma cell line (Raji).

作者信息

Gao Jianming, Luo Xiaomin, Tang Ke, Li Xiaoling, Li Guiyuan

机构信息

Cancer Research Institute, Xiang-Ya School of Medicine, Central South University, Changsha, Hunan 410078, China.

出版信息

J Virol Methods. 2006 Sep;136(1-2):193-9. doi: 10.1016/j.jviromet.2006.05.013. Epub 2006 Jun 23.

Abstract

Epstein-Barr virus (EBV) integration into a Burkitt's lymphoma (BL) cell line (Raji) was investigated, using polymerase chain reaction (PCR), Southern hybridization, genomic library screening and fluorescence in situ hybridization (FISH). BaMHIW fragments of the EBV genome and DNA sequences of the viral latent membrane protein (LMP)1 and LMP2 genes were detected in Raji cells. BaMHI-digested high-molecular weight DNA from Raji cells generated 4 and 10 kb, 23 kb fragments that hybridized to Probe-1 (EBV genome 13232-16189) and Probe-2 (EBV genome 5-3271). Genomic library for Raji cells was constructed. Plaques (1 x 10(5)) were screened with Probe-2, and four positive clones were obtained. Chromosomal integration of EBV DNA was detected in the Raji cell. The viral integration sites included 1p, 1q, 2q, 3p, 3q, 4q, 5q, 6q, 7p, 7q, 9q, 11p, 14q and 15q. Despite this multiplicity of integration sites, integration showed high frequency only at the sites 4q, 2q, 1q and 7q; 64% of the total signals were found in these four chromosomal bands. No viral integration occurred in chromosomes 16-22 or the sex chromosomes (X, Y). This study is the first comprehensive FISH analysis of EBV integration into the chromosomes of the Raji cell line. The findings support the notion that EBV integrates into the Raji cell genome non-randomly.

摘要

采用聚合酶链反应(PCR)、Southern杂交、基因组文库筛选及荧光原位杂交(FISH)技术,对爱泼斯坦-巴尔病毒(EBV)整合进伯基特淋巴瘤(BL)细胞系(Raji)的情况进行了研究。在Raji细胞中检测到了EBV基因组的BaMHIW片段以及病毒潜伏膜蛋白(LMP)1和LMP2基因的DNA序列。来自Raji细胞经BaMHI酶切的高分子量DNA产生了4 kb和10 kb、23 kb的片段,这些片段与探针1(EBV基因组13232 - 16189)和探针2(EBV基因组5 - 3271)杂交。构建了Raji细胞的基因组文库。用探针2筛选了1×10⁵个噬菌斑,获得了4个阳性克隆。在Raji细胞中检测到了EBV DNA的染色体整合。病毒整合位点包括1p、1q、2q、3p、3q、4q、5q、6q、7p、7q、9q、11p、14q和15q。尽管整合位点具有多样性,但仅在4q、2q、1q和7q位点出现了高频整合;在这4条染色体带中发现了64%的总信号。在16 - 22号染色体或性染色体(X、Y)上未发生病毒整合。本研究是对EBV整合进Raji细胞系染色体的首次全面FISH分析。研究结果支持EBV非随机整合进Raji细胞基因组这一观点。

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