Zink R T, Engwall J K, McEvoy J L, Chatterjee A K
J Bacteriol. 1985 Oct;164(1):390-6. doi: 10.1128/jb.164.1.390-396.1985.
Pectin lyase (PNL) and the bacteriocin carotovoricin (CTV) were induced in Erwinia carotovora subsp. carotovora 71 by the DNA-damaging agents mitomycin C, nalidixic acid, and UV light. To determine whether the recA product was involved in the expression of these damage-inducible phenotypes, we cloned the E. carotovora subsp. carotovora recA+ gene, inactivated it by Tn5 insertion, and constructed an E. carotovora subsp. carotovora recA::Tn5 strain by gene replacement via homologous recombination. The RecA- strain was more sensitive to methyl methanesulfonate, nitroquinoline oxide, and UV light than its RecA+ parent. The recA mutation did not affect the production of pectate lyase, polygalacturonase, cellulase, and protease or the ability to cause soft rot of potato tubers. With this mutant, unlike with the RecA+ parent strain, PNL and CTV were not induced by mitomycin C or detected in potato tuber tissue. The RecA+ phenotype, including the inducibility of PNL and CTV, could, however, be restored in the mutant in trans by the recA+ gene from either E. carotovora subsp. carotovora or Escherichia coli. We conclude that, in E. carotovora subsp. carotovora, the recA product is required in the induction of PNL and CTV.
在胡萝卜软腐欧文氏菌胡萝卜软腐亚种71中,DNA损伤剂丝裂霉素C、萘啶酸和紫外线可诱导果胶裂解酶(PNL)和细菌素胡萝卜软腐欧文氏菌素(CTV)的产生。为了确定recA基因产物是否参与这些损伤诱导型表型的表达,我们克隆了胡萝卜软腐欧文氏菌胡萝卜软腐亚种的recA⁺基因,通过Tn5插入使其失活,并通过同源重组进行基因替换构建了胡萝卜软腐欧文氏菌胡萝卜软腐亚种recA::Tn5菌株。RecA⁻菌株对甲磺酸甲酯、氧化硝基喹啉和紫外线比其RecA⁺亲本更敏感。recA突变不影响果胶酸裂解酶、聚半乳糖醛酸酶、纤维素酶和蛋白酶的产生,也不影响引起马铃薯块茎软腐的能力。与RecA⁺亲本菌株不同,对于该突变体,丝裂霉素C不会诱导PNL和CTV的产生,在马铃薯块茎组织中也检测不到它们。然而,来自胡萝卜软腐欧文氏菌胡萝卜软腐亚种或大肠杆菌的recA⁺基因可以在反式条件下使该突变体恢复RecA⁺表型,包括PNL和CTV的可诱导性。我们得出结论,在胡萝卜软腐欧文氏菌胡萝卜软腐亚种中,recA基因产物是诱导PNL和CTV所必需的。