DIFFERENTIAL SUBSEQUENCE CONSERVATION OF INTERSPERSED REPETITIVE STREPTOCOCCUS-PNEUMONIAE BOX ELEMENTS IN DIVERSE BACTERIA
DIFFERENTIAL SUBSEQUENCE CONSERVATION OF INTERSPERSED REPETITIVE STREPTOCOCCUS-PNEUMONIAE BOX ELEMENTS IN DIVERSE BACTERIA
复制标题
DOI：
10.1101/gr.5.4.408
复制
发表时间：
1995-11-01
期刊：
影响因子：
7
通讯作者：
LUPSKI, JR
中科院分区：
文献类型：
--
作者：
KOEUTH, T;VERSALOVIC, J;LUPSKI, JR
研究了肺炎链球菌穿插重复DNA序列BOX的进化保守性，以探索这些元件的镶嵌性质。Box元素由BoxA、BoxB和BoxC三个子单元的各种组合组成。根据BoxA、BoxB和BoxC亚基的一致DNA序列设计了8个寡核苷酸探针。DNA杂交研究和使用这些探针/引物的聚合酶链式反应表明，BoxA亚单位内的寡核苷酸序列在不同的细菌物种中似乎是保守的。BoxB和boxC亚基在肺炎链球菌以外的细菌中仅显示有限的序列保守性(如果有的话)。含有BoxA、BoxB和BoxC亚基的完整盒元件仅在肺炎球菌株中以高拷贝数存在。这种差异保守模式支持了box重复元件的模块化性质，因为在肺炎链球菌以外的细菌中，boxA样亚序列有效地独立于boxB或boxC样亚单位。此外，使用BOXA1R引物从肺炎链球菌分离株的基于重复序列的聚合酶链式反应(REP-PCR)指纹中获得的树状图产生了与以前通过其他方法获得的相似的聚类图，这表明这些基于重复序列的DNA指纹代表了肺炎链球菌基因组的内在特征。我们的结果表明在细菌界中广泛存在BoxA样亚序列的保守，支持肺炎链球菌中Box的嵌合性，并证明了基于BoxA的引物在许多微生物的rep-PCR指纹图谱中的实用性。
Evolutionary conservation of an interspersed repetitive DNA sequence, BOX, from Streptococcus pneumoniae was investigated to explore the mosaic nature of these elements. BOX elements consist of various combinations of three subunits, boxA, boxB, and boxC. Eight oligonucleotide probes were designed based on consensus DNA sequences of boxA, boxB, and boxC subunits. DNA hybridization studies and PCR using these probes/primers demonstrate that oligonucleotide sequences within the boxA subunit appear to be conserved among diverse bacterial species. The boxB and boxC subunits show only limited, if any, sequence conservation in bacteria other than S. pneumoniae. Intact BOX elements with boxA, boxB, and boxC subunits were only present in high copy number in pneumococcal strains. This pattern of differential conservation lends support to the modular nature of BOX repetitive elements in that boxA-like subsequences are effectively independent of boxB-like or boxC-like subunits in bacteria other than S. pneumoniae. Furthermore, dendrograms derived from repetitive sequence-based PCR (rep-PCR) fingerprints of S. pneumoniae isolates using the BOXA1R primer yielded clustering patterns that were similar to those obtained previously by other methods, suggesting that these repetitive sequence-based DNA fingerprints represent intrinsic properties of an S. pneumoniae strain's genome. Our results indicate widespread conservation of boxA-like subsequences in the bacterial kingdom, lend support to the mosaic nature of BOX in S. pneumoniae, and demonstrate the utility of boxA-based primers for rep-PCR fingerprinting of many microorganisms.