Zyskind J W, Smith D W
Proc Natl Acad Sci U S A. 1980 May;77(5):2460-4. doi: 10.1073/pnas.77.5.2460.
Construction of deletion derivative plasmids and cloning of restriction fragments from plasmids containing the Salmonella typhimurium origin of replication (ori) were used to locate the functional origin to within a DNA fragment of 296 base pairs between the genes uncB and asn. The nucleotide sequence of the S. typhimurium ori region was determined and compared with the Escherichia coli ori sequence. In the 296-base pair fragment, 85.8% of the bases are conserved between the two species. A nearly equal number of transition and transversion type differences, with no insertions or deletions, occurs between the two bacterial origins, such that the relatively high percentage (adenine plus thymine) of 59.5% is conserved. The 296-base pair fragment contains 14 GATC sequences, all of which are conserved. The high frequency of occurrence of GATC, which is the site of methylation under control of the dam gene, may explain in part why the bacterial ori region appears to be so highly conserved. A large number of secondary structures are possible. One such structure, with a "cloverleaf," is favored by ori nucleotide sequence comparisons and leads to potential novel macromolecular interactions.
构建缺失衍生质粒并从含有鼠伤寒沙门氏菌复制起点(ori)的质粒中克隆限制片段,用于将功能起点定位在uncB和asn基因之间296个碱基对的DNA片段内。测定了鼠伤寒沙门氏菌ori区域的核苷酸序列,并与大肠杆菌ori序列进行了比较。在296个碱基对的片段中,两个物种之间85.8%的碱基是保守的。两个细菌起点之间出现了数量几乎相等的转换和颠换类型差异,没有插入或缺失,因此59.5%的相对高比例(腺嘌呤加胸腺嘧啶)得以保守。296个碱基对的片段包含14个GATC序列,所有这些序列都是保守的。GATC的高出现频率是dam基因控制下的甲基化位点,这可能部分解释了为什么细菌ori区域似乎高度保守。可能存在大量二级结构。一种具有“三叶草”的结构,受到ori核苷酸序列比较的青睐,并导致潜在的新型大分子相互作用。