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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >SITE-SPECIFIC NICKING IN VITRO AT ORI T BY THE DNA RELAXASE OF TN5252
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SITE-SPECIFIC NICKING IN VITRO AT ORI T BY THE DNA RELAXASE OF TN5252

机译：TN5252的DNA松弛酶在体外进行特异性位点定位

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摘要

Tn5252 is a promiscuous streptococcal element capable of madiating horizontal spread of multiple antibiotic resistance. To begin understanding the functional role of a transfer-related region in Tn5252, its nucleotide sequence was determined. Sequence of this 3.3-kb DNA segment revealed the presence of six open reading frames. The predicted amino acid sequence of one of the open reading frames, ORF9, showed similarity to a predicted protein product of the lactococcal conjugative plasmid, pC1528. The deduced primary protein sequence of another, ORF4, showed strong structural similarity to conserved regions of various prokaryotic DNA relaxases that initiate conjugal transfer by strand- and site-specific cleavage at the transfer origin. A hybrid protein containing the ORF4 protein fused to the carboxyl terminal end of maltose binding protein was purified from Escherichia coli and found to specifically nick plasmids carrying a 2-kb DNA segment derived from the transposon. The nicking reaction is protein concentration-dependent. These results imply that the conjugative transposition of Tn5252 may involve rolling circle replication and transfer of a unique DNA strand. (C) 1997 Academic Press. [References: 25]
机译：Tn5252是一种混杂的链球菌元件，能够缓解多种抗生素耐药性的水平扩散。为了开始了解Tn5252中转移相关区域的功能，确定了其核苷酸序列。该3.3kb DNA片段的序列揭示了六个开放阅读框的存在。开放阅读框之一ORF9的预测氨基酸序列显示与乳球菌结合质粒pC1528的预测蛋白质产物相似。推导的另一个蛋白ORF4的一级蛋白质序列显示出与各种原核DNA松弛酶保守区域的强结构相似性，这些原核DNA松弛酶通过在转移起点的链和位点特异性裂解来启动夫妻转移。从大肠杆菌中纯化了含有融合至麦芽糖结合蛋白的羧基末端的ORF4蛋白的杂合蛋白，并发现其特异地刻有携带来自转座子的2-kb DNA片段的质粒。切口反应是蛋白质浓度依赖性的。这些结果暗示Tn5252的共轭转座可能涉及滚环复制和独特DNA链的转移。 （C）1997学术出版社。 [参考：25]

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