MOLECULAR-CLONING OF THE COMPLETE EPSTEIN-BARR VIRUS GENOME AS A SET OF OVERLAPPING RESTRICTION ENDONUCLEASE FRAGMENTS
MOLECULAR-CLONING OF THE COMPLETE EPSTEIN-BARR VIRUS GENOME AS A SET OF OVERLAPPING RESTRICTION ENDONUCLEASE FRAGMENTS
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DOI：
10.1093/nar/9.13.2999
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发表时间：
1981-01-01
影响因子：
14.9
通讯作者：
GRIFFIN, BE
中科院分区：
文献类型：
--
作者：
ARRAND, JR;RYMO, L;GRIFFIN, BE
用限制性内切酶ecori切割获得的爱泼斯坦-巴尔病毒DNA的完整片段已被克隆。克隆了来自B95-8株线性病毒粒子DNA的14个不同的病毒基因组内部片段，以及来自B95-8细胞内圆形EBV DNA的病毒粒子DNA末端区域对应的序列。通过琼脂糖凝胶电泳测定了片段的大小，它们的总和导致病毒粒子DNA的分子量估计为110 × 106。利用另一种EBV DNA来源，即来自Raji细胞的环状病毒DNA，也在cosmids中克隆了特别感兴趣的大型ecori DNA片段。为了提供一组重叠序列，在pBR322中克隆了B95-8病毒粒子dna的所有29个内部bamhi片段。每个克隆的DNA片段在病毒基因组中的定位是通过与几种不同的限制性内切酶切割的病毒粒子DNA杂交来确定的。
A complete collection of fragments of Epstein-Barr virus DNA, obtained by cleavage with restriction endonucleaseEcoRI, has been cloned. Fourteen different internal fragments of the virus genome, derived from linear virion DNA of the B95-8 strain, and sequences corresponding to the terminal regions of virion DNA, derived from intracel1ular circular EBV DNA isolated from B95-8 cells, were cloned. Sizes of fragments were determined by agarose gel electrophoresis and their sum leads to an estimated molecular weight of 110 × 106for virion DNA. LargeEcoRI DNA fragments of special interest were also cloned in cosmids using another source of EBV DNA, that is, the circular viral DNA derived from Raji cells. In order to provide a set of overlapping sequences, all the 29 internalBamHI fragments of B95-8 virion DNAwere cloned in pBR322. The map location with in the viral genome of each cloned DNA fragment was identified by hybridizing to blots of virion DNA cleaved with several different restriction endonucleases.