Wiedmann M, Arvik T J, Hurley R J, Boor K J
Department of Food Science, College of Agriculture and Life Sciences, Cornell University, Ithaca, New York 14853, USA.
J Bacteriol. 1998 Jul;180(14):3650-6. doi: 10.1128/JB.180.14.3650-3656.1998.
The gene encoding the general stress transcription factor sigmaB in the gram-positive bacterium Listeria monocytogenes was isolated with degenerate PCR primers followed by inverse PCR amplification. Evidence for gene identification includes the following: (i) phylogenetic analyses of reported amino acid sequences for sigmaB and the closely related sigmaF proteins grouped L. monocytogenes sigmaB in the same cluster with the sigmaB proteins from Bacillus subtilis and Staphylococcus aureus, (ii) the gene order in the 2, 668-bp portion of the L. monocytogenes sigB operon is rsbU-rsbV-rsbW-sigB-rsbX and is therefore identical to the order of the last five genes of the B. subtilis sigB operon, and (iii) an L. monocytogenes sigmaB mutant had reduced resistance to acid stress in comparison with its isogenic parent strain. The sigB mutant was further characterized in mouse models of listeriosis by determining recovery rates of the wild-type and mutant strains from livers and spleens following intragastric or intraperitoneal infection. Our results suggest that sigmaB-directed genes do not appear to be essential for the spread of L. monocytogenes to mouse liver or spleen at 2 and 4 days following intragastric or intraperitoneal infection.
利用简并PCR引物分离出革兰氏阳性菌单核细胞增生李斯特菌中编码一般应激转录因子σB的基因,随后进行反向PCR扩增。基因鉴定的证据如下:(i) 对已报道的σB和密切相关的σF蛋白氨基酸序列进行系统发育分析,结果表明单核细胞增生李斯特菌的σB与枯草芽孢杆菌和金黄色葡萄球菌的σB蛋白归为同一簇;(ii) 单核细胞增生李斯特菌sigB操纵子2668 bp部分的基因顺序为rsbU-rsbV-rsbW-sigB-rsbX,因此与枯草芽孢杆菌sigB操纵子最后五个基因的顺序相同;(iii) 与同基因亲本菌株相比,单核细胞增生李斯特菌σB突变体对酸应激的抗性降低。通过测定野生型和突变体菌株在胃内或腹腔感染后从肝脏和脾脏中的回收率,在李斯特菌病小鼠模型中对sigB突变体进行了进一步表征。我们的结果表明,在胃内或腹腔感染后2天和4天时,σB指导的基因似乎对单核细胞增生李斯特菌向小鼠肝脏或脾脏的传播不是必需的。