Rasmussen O F, Beck T, Olsen J E, Dons L, Rossen L
Department of Molecular Food Technology, Biotechnological Institute, Lyngby, Denmark.
Infect Immun. 1991 Nov;59(11):3945-51. doi: 10.1128/iai.59.11.3945-3951.1991.
The nucleotide sequence of a 3.5-kb BamHI fragment from Listeria monocytogenes 12067, a human clinical isolate of serotype 4b, has been determined. The DNA fragment harbors the gene for listeriolysin, part of the gene for a phosphatidylinositol-specific phospholipase C, and part of the gene for a metalloprotease. Comparison of the sequence with corresponding sequences from two other L. monocytogenes isolates revealed a significant number of nucleotide differences. Several of the differences give rise to amino acid substitutions. The most variable region was the examined part of the mpl gene, whereas the lisA gene showed a relatively high degree of conservation, particularly at the amino acid level. To analyze the pattern of sequence variability in the lisA gene, a 160-bp region covering nine nucleotide differences was sequenced from 36 isolates of different origins. This work showed that the strains can be grouped into two major types according to the nucleotide sequences. Oligonucleotide probing of a larger number of L. monocytogenes isolates showed that the observed differences can be used to subdivide the species. The data suggest a correspondence between the sequence type of the lisA gene and flagellar antigens. Assays based on hybridization or the polymerase chain reaction with type-specific oligonucleotides may provide fast and easy alternative methods for strain typing.
已确定了来自产单核细胞李斯特菌12067(血清型4b的人类临床分离株)的一个3.5kb BamHI片段的核苷酸序列。该DNA片段含有李斯特菌溶血素基因、磷脂酰肌醇特异性磷脂酶C基因的一部分以及金属蛋白酶基因的一部分。将该序列与另外两个产单核细胞李斯特菌分离株的相应序列进行比较,发现了大量的核苷酸差异。其中一些差异导致了氨基酸替换。变化最大的区域是mpl基因的检测部分,而lisA基因显示出相对较高的保守程度,尤其是在氨基酸水平。为了分析lisA基因的序列变异模式,从36个不同来源的分离株中对覆盖9个核苷酸差异的160bp区域进行了测序。这项工作表明,根据核苷酸序列,这些菌株可分为两种主要类型。对大量产单核细胞李斯特菌分离株进行寡核苷酸探测表明,观察到的差异可用于对该菌种进行细分。数据表明lisA基因的序列类型与鞭毛抗原之间存在对应关系。基于杂交或使用型特异性寡核苷酸的聚合酶链反应的检测方法可能为菌株分型提供快速简便的替代方法。