SEQUENCE SPECIFICITY OF BACILLUS-SUBTILIS DNA GYRASE INVIVO
SEQUENCE SPECIFICITY OF BACILLUS-SUBTILIS DNA GYRASE INVIVO
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DOI：
10.1007/bf00056101
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发表时间：
1991-01-01
期刊：
影响因子：
1.5
通讯作者：
ZVINGILA, DJ
中科院分区：
文献类型：
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作者：
BASHKIROV, VI;ZVINGILA, DJ
通过用十二烷基硫酸钠裂解恶喹酸处理的枯草芽孢杆菌原生质体，对pBG 0（大肠杆菌质粒pBR 322和金黄色葡萄球菌质粒pUB 110之间的突变体）进行线性化。该质粒DNA线性化既用于各种强度的DNA促旋酶切割位点的详细作图，又用于通过引入XhoI接头DNA在促旋酶介导的切割点处的核苷酸序列测定。通过标记XhoI位点的3'末端，对40个带有XhoI接头的质粒进行了测序，发现其中38个质粒在接头两侧含有4个碱基对的重复序列，这是恶喹酸诱导E. coli DNA旋转酶的体外和体内研究。这些测序位点的相对强度通过将它们的位置与定位在适当质粒基因组上的位点进行比较来确定。这使我们能够提出B的共有序列。枯草杆菌DNA旋转酶体内切割位点：GNAT\ GATCATNC。(T)其中N是任何核苷酸。括号中的碱是次要的。DNA旋转酶参与枯草芽孢杆菌的非法重组事件进行了讨论。
Linearization of pBG0 (a hydrid between Escherichia coli plasmid pBR322 and Staphylococcus aureus plasmid pUB110) was performed by lysis of the oxolinic acid treated Bacillus subtilis protoplasts with sodium dodecyl sulfate. This plasmid DNA linearization was used both for a detailed mapping of DNA gyrase cleavage sites of various strength and for the nucleotide sequence determinations at the points of gyrase-mediated scissions by introducing the XhoI linker DNA. A total of 40 plasmids carrying inserted XhoI linker were sequenced by labeling 3' termini of XhoI sites; 38 of them were found to contain a duplication of four base-pairs of the plasmid sequence flanking the linker, which were characteristic of the oxolinic acid-induced DNA cleavage by E. coli DNA gyrase in vitro and in vivo. The relative strength of these sequenced sites was established by comparing their positions to the sites mapped on the appropriate plasmid genome. This allowed us to propose a consensus sequence of B. subtilis DNA gyrase in vivo cleavage site: GNAT\ GATCATNC. (T) (G) (A) where N is any nucleotide. The bases in parentheses were preferred secondarily. The involvement of DNA gyrase in illegitimate recombination events in Bacillus subtilis is discussed.