Construction of a cloned library of the EcoRI fragments from the human cytomegalovirus genome (strain AD169)
Construction of a cloned library of the EcoRI fragments from the human cytomegalovirus genome (strain AD169)
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DOI：
10.1128/jvi.42.2.547-557.1982
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发表时间：
1982-05
影响因子：
5.4
通讯作者：
J. C. Tamashiro;L. Hock;D. Spector
中科院分区：
文献类型：
--
作者：
J. C. Tamashiro;L. Hock;D. Spector
人巨细胞病毒（HCMV）AD 169株的DNA基因组为158 × 10（6）Mr。用限制性内切酶EcoRI切割HCMV DNA产生35个大小在0.54 × 10（6）Mr范围内的主要片段。使用质粒pACYC 184和受体细菌大肠杆菌菌株HB 101 RecA-。通过与病毒DNA杂交来确定克隆插入片段的病毒来源。通过用其他限制性内切酶消化进一步表征片段。获得了几个克隆，其含有跨越病毒DNA序列的长（L）和短（S）组分之间的连接的序列。这些克隆的分子量相差0.3 x 10（6）至0.4 x 10（6）Mr的倍数。在克隆中发现的变异性也反映在基因组中。每个克隆含有一个连接序列，与EcoRI消化的HCMV DNA Southern滤膜上的一系列条带杂交。这种“阶梯效应”为L-S连接处的异质性区域提供了证据。
The DNA genome of human cytomegalovirus (HCMV) strain AD169 is 158 x 10(6) Mr. Cleavage of the HCMV DNA with the restriction endonuclease EcoRI yields 35 major fragments ranging in size from 0.54 x 10(6) Mr. We have constructed a cloned library of the EcoRI fragments of this strain of HCMV, using the plasmid pACYC184 and the recipient bacterium Escherichia coli strain HB101 RecA-. The viral origin of the cloned inserts was determined by hybridization to viral DNA. The fragments were characterized further by digestion with other restriction enzymes. Several clones were obtained which contained sequences spanning the junction between the long (L) and short (S) components of the viral DNA sequences. These clones differed in molecular weight by multiples of 0.3 x 10(6) to 0.4 x 10(6) Mr. The variability found in the clones was also reflected in the genome. Each clone containing a junction sequence hybridized to a series of bands on Southern filters of EcoRI-digested HCMV DNA. This "ladder effect" provided evidence for a region of heterogeneity within the L-S junction.