Stibitz S, Yang M S
Division of Bacterial Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA.
J Bacteriol. 1997 Sep;179(18):5820-6. doi: 10.1128/jb.179.18.5820-5826.1997.
The genomic organization of Bordetella pertussis strains has been examined by using a new method. This method does not depend on the prior determination of a restriction map of the bacterial chromosome but is based on the ability to measure directly the distance between two genes. This is accomplished through the integration at each gene of a suicide vector containing a cleavage site for the intron-encoded endonuclease I-SceI, which is not otherwise found in the chromosome. Integration is mediated by homologous recombination between the chromosomal and cloned plasmid copies of a gene of interest. Digestion with I-SceI gives rise to a fragment the size of which represents the distance between the two genes. Multiple pairwise determinations within a set of genes provide sufficient information to derive a map of the relative gene positions. Mapping a set of 11 to 13 genes for five strains of B. pertussis and one strain of B. parapertussis revealed extensive divergence of gene order between B. pertussis Tohama I, B. pertussis 18-323, and B. parapertussis ATCC 15311. Less extensive divergence of gene order was observed between B. pertussis Tohama I and B. pertussis Tohama III, BP165, and Wellcome 28, with most of the observed differences explainable by large inversions.
已使用一种新方法对百日咳博德特氏菌菌株的基因组组织进行了研究。该方法不依赖于事先确定细菌染色体的限制性图谱，而是基于直接测量两个基因之间距离的能力。这是通过在每个基因处整合一个自杀载体来实现的，该载体含有内含子编码的内切核酸酶I-SceI的切割位点，而该位点在染色体中其他地方不存在。整合由感兴趣基因的染色体拷贝和克隆质粒拷贝之间的同源重组介导。用I-SceI消化会产生一个片段，其大小代表两个基因之间的距离。一组基因内的多个成对测定提供了足够的信息来推导相对基因位置的图谱。对五株百日咳博德特氏菌和一株副百日咳博德特氏菌的11至13个基因进行图谱绘制，结果显示百日咳博德特氏菌Tohama I、百日咳博德特氏菌18-323和副百日咳博德特氏菌ATCC 15311之间的基因顺序存在广泛差异。在百日咳博德特氏菌Tohama I与百日咳博德特氏菌Tohama III、BP165和Wellcome 28之间观察到的基因顺序差异较小，观察到的大多数差异可用大的倒位来解释。
