Ito H, Kido N, Arakawa Y, Ohta M, Sugiyama T, Kato N
Department of Bacteriology, Nagoya University School of Medicine, Aichi, Japan.
Appl Environ Microbiol. 1991 Oct;57(10):2912-7. doi: 10.1128/aem.57.10.2912-2917.1991.
A Southern hybridization analysis revealed that the region homologous to Escherichia coli lacZ was present on the chromosomal DNAs of beta-galactosidase-positive Shigella strains, such as Shigella dysenteriae serovar 1 and Shigella sonnei strains, whereas this region was absent from chromosomal DNAs of beta-galactosidase-negative strains of Shigella flexneri and Shigella boydii. We found that the lacY-A region was deficient in S. dysenteriae serovar 1 and believe that this is the reason for the slow fermentation of lactose by this strain. S. sonnei strains possessed the region which hybridized with E. coli lacY-A despite their slow hydrolysis of lactose. The whole lactose-fermenting region was cloned from S. sonnei and compared with the cloned lac operon of E. coli K-12. Both clones directed the synthesis of beta-galactosidase in an E. coli K-12 strain lacking indigenous beta-galactosidase activity (strain JM109-1), and we observed no difference in the expression of beta-galactosidase activity in S. sonnei and E. coli. However, E. coli JM109-1 harboring the lactose-fermenting genes of S. sonnei exhibited the slow lactose fermentation phenotype like the parental strain. S. sonnei strains had no detectable lactose permease activities. E. coli JM109-1 harboring the lactose-fermenting genes of S. sonnei had a detectable permease activity, possibly because of the multicopy nature of the cloned genes, but this permease activity was much lower than that of strain JM109-1 harboring the lac operon of E. coli K-12. From these results we concluded that slow lactose fermentation by S. sonnei is due to weak lactose permease activity.
Southern杂交分析显示,与大肠杆菌lacZ同源的区域存在于β-半乳糖苷酶阳性志贺氏菌菌株的染色体DNA上,如痢疾志贺氏菌血清型1和宋内志贺氏菌菌株,而在弗氏志贺氏菌和鲍氏志贺氏菌的β-半乳糖苷酶阴性菌株的染色体DNA中不存在该区域。我们发现痢疾志贺氏菌血清型1中lacY - A区域缺失,并且认为这是该菌株乳糖发酵缓慢的原因。宋内志贺氏菌菌株尽管乳糖水解缓慢,但拥有与大肠杆菌lacY - A杂交的区域。从宋内志贺氏菌中克隆了整个乳糖发酵区域,并与克隆的大肠杆菌K - 12的lac操纵子进行比较。这两个克隆在缺乏自身β-半乳糖苷酶活性的大肠杆菌K - 12菌株(JM109 - 1菌株)中都指导了β-半乳糖苷酶的合成,并且我们观察到宋内志贺氏菌和大肠杆菌中β-半乳糖苷酶活性的表达没有差异。然而,携带宋内志贺氏菌乳糖发酵基因的大肠杆菌JM109 - 1表现出与亲代菌株相似的缓慢乳糖发酵表型。宋内志贺氏菌菌株没有可检测到的乳糖通透酶活性。携带宋内志贺氏菌乳糖发酵基因的大肠杆菌JM109 - 1具有可检测到的通透酶活性,这可能是由于克隆基因的多拷贝性质,但这种通透酶活性远低于携带大肠杆菌K - 12的lac操纵子的JM109 - 1菌株。从这些结果我们得出结论,宋内志贺氏菌乳糖发酵缓慢是由于乳糖通透酶活性较弱。