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The gene coding for protein p60 of Listeria monocytogenes and its use as a specific probe for Listeria monocytogenes.

机译:编码单核细胞增生李斯特氏菌蛋白p60的基因及其用作单核细胞增生李斯特菌的特异性探针。

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摘要

The gene of Listeria monocytogenes that encodes a major extracellular protein (p60) was cloned in Escherichia coli. The gene was designated iap, as p60 was previously shown to represent an invasion-associated protein (M. Kuhn and W. Goebel, Infect. Immun. 57:55-61, 1989). The recombinant E. coli clone expressed p60, as shown by immunoblotting. The complete nucleotide sequence of iap was determined. The deduced amino acid sequence of p60 (484 amino acids) contains a putative N-terminal signal sequence of 27 amino acids and an extended repeat region consisting of 19 threonine-asparagine units. Hybridization with the entire iap gene revealed the presence of homologous sequences in most other Listeria species. In contrast, a 400-base-pair internal iap probe which contained the whole repeat region hybridized only with genomic DNA from L. monocytogenes. Four oligonucleotides previously described as specific probes for the detection of L. monocytogenes (A. R. Datta, B. A. Wentz, D. Shook, and M. W. Trucksess, Appl. Environ. Microbiol. 54:2933-2937, 1988) were shown to be part of the iap gene.
机译:在大肠杆菌中克隆了编码主要细胞外蛋白(p60)的单核细胞增生李斯特菌基因。该基因被命名为Iap,因为先前显示p60代表入侵相关蛋白(M. Kuhn and W. Goebel,Infect。Immun。57:55-61,1989)。如免疫印迹所示,重组大肠杆菌克隆表达了p60。确定了iap的完整核苷酸序列。推导的p60氨基酸序列(484个氨基酸)包含一个推定的27个氨基酸的N末端信号序列和一个由19个苏氨酸-天冬酰胺单元组成的延伸重复区域。与整个iap基因的杂交揭示了在大多数其他李斯特菌属物种中存在同源序列。相反,包含整个重复区的400个碱基对的内部Iap探针仅与单核细胞增生李斯特氏菌的基因组DNA杂交。显示了先前描述为检测单核细胞增生李斯特氏菌的特异性探针的四种寡核苷酸(AR Datta,BA Wentz,D.Shook和MW Trucksess,Appl.Environ.Microbiol.54:2933-2937,1988)。 IAP基因。

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