Schmidt H, Scheef J, Janetzki-Mittmann C, Datz M, Karch H
Institut für Hygiene und Mikrobiologie, Universität Würzburg, Germany.
FEMS Microbiol Lett. 1997 Apr 1;149(1):39-44. doi: 10.1111/j.1574-6968.1997.tb10305.x.
A cosmid library of enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain EDL 933 was constructed and clones carrying the stx2 gene were identified by colony blot hybridization with a stx2B specific probe. Nucleotide sequencing upstream of the stx2A gene revealed high sequence identities of 89.5% to the ileX tRNA gene found in E. coli. The ileX gene was located 260 bp from the translational start codon of stx2A. PCR analysis with primers specific for this analyzed region showed that in 11 Stx2-producing EHEC strains from patients with hemolytic uremic syndrome, all PCR-positive strains carried the ileX tRNA gene. However, PCR analysis of the respective region in 11 Stxl-producing EHEC strains detected no ileX genes. Although the role of ileX in Stx2-producing EHEC strains is not clear, its function in regard to the use of rare codons and as an integration site is discussed.
构建了肠出血性大肠杆菌(EHEC)O157:H7菌株EDL 933的黏粒文库,并通过用stx2B特异性探针进行菌落杂交鉴定携带stx2基因的克隆。stx2A基因上游的核苷酸测序显示与大肠杆菌中发现的ileX tRNA基因具有89.5%的高序列同一性。ileX基因位于stx2A翻译起始密码子260 bp处。用针对该分析区域的特异性引物进行PCR分析表明,在11株来自溶血尿毒综合征患者的产Stx2的EHEC菌株中,所有PCR阳性菌株都携带ileX tRNA基因。然而,对11株产Stx1的EHEC菌株的相应区域进行PCR分析未检测到ileX基因。尽管ileX在产Stx2的EHEC菌株中的作用尚不清楚,但讨论了其在稀有密码子使用方面的功能以及作为整合位点的功能。