Zyskind J W, Cleary J M, Brusilow W S, Harding N E, Smith D W
Proc Natl Acad Sci U S A. 1983 Mar;80(5):1164-8. doi: 10.1073/pnas.80.5.1164.
The chromosomal replication origin (oriC) of Vibrio harveyi has been isolated on a plasmid and shown to function as an origin in Escherichia coli. The nucleotide sequence of the V. harveyi oriC was determined. From a comparison of this sequence with oriC sequences of five enteric bacteria, we derived a consensus sequence of bacterial origins that function in E. coli. This consensus sequence identifies 122 positions within oriC where nucleotide substitutions can occur without loss of origin function. These positions are clustered rather than scattered. Four interrelated nine-base-pair repeats and eight of the dam methylation G-A-T-C sites are conserved in the consensus sequence. Very few relative insertion-deletion changes occur, and these are localized to one region of oriC. The genes for three polypeptides linked to the V. harveyi oriC were identified by using in vitro protein synthesis directed by deletion derivative plasmid templates. One of these genes, coding for a 58,000 Mr polypeptide and located 3.0 kilobase pairs from the V. harveyi oriC region, is lethal to E. coli when many copies (approximately 40 per cell) are present (high copy lethal or HCL gene). In addition, nucleotide sequence analysis showed that a different gene, the gid gene to the left of oriC, is highly conserved between E. coli and V. harveyi, whereas the coding region to the right of oriC is much less conserved.
哈维氏弧菌的染色体复制起点（oriC）已在质粒上分离出来，并证明其在大肠杆菌中可作为复制起点发挥作用。已测定了哈维氏弧菌oriC的核苷酸序列。通过将该序列与五种肠道细菌的oriC序列进行比较，我们得出了在大肠杆菌中起作用的细菌复制起点的共有序列。该共有序列确定了oriC内122个位置，在这些位置发生核苷酸替换不会丧失复制起点功能。这些位置是成簇的而非分散的。四个相互关联的九碱基对重复序列和八个dam甲基化G - A - T - C位点在共有序列中是保守的。相对的插入 - 缺失变化很少发生，并且这些变化局限于oriC的一个区域。通过使用由缺失衍生物质粒模板指导的体外蛋白质合成，鉴定了与哈维氏弧菌oriC相关的三种多肽的基因。其中一个基因编码一种58,000 Mr的多肽，位于距哈维氏弧菌oriC区域3.0千碱基对处，当存在许多拷贝（每个细胞约40个）时对大肠杆菌是致死的（高拷贝致死或HCL基因）。此外，核苷酸序列分析表明，oriC左侧的一个不同基因，即gid基因，在大肠杆菌和哈维氏弧菌之间高度保守，而oriC右侧的编码区域保守性则低得多。
