NaClO脅迫下單核細胞增生李斯特菌WaX12及其sigB基因缺失突變株抗氧化脅迫相關(guān)功能基因表達比較
發(fā)布時間:2018-05-03 22:44
本文選題:單增李斯特菌 + sigB; 參考:《食品科學》2017年10期
【摘要】:目的:探究單核細胞增生李斯特菌sigB基因?qū)Υ温人徕c(NaClO)脅迫的調(diào)控作用。方法:在半致死濃度NaClO(3 mmol/L)脅迫下,比較該菌野生株WaX12與sigB基因敲除突變株的耐受程度及菌內(nèi)的活性氧水平,并通過實時定量聚合酶鏈式反應技術(shù)(real-time quantitative polymerase chain reaction,RT-PCR)進一步觀察lmo1433、lmo0906、lmo2344、ohrR、lmo2770基因隨脅迫時間的轉(zhuǎn)錄水平變化。結(jié)果:野生株和sigB突變株的表型測定結(jié)果顯示,野生株比缺失株更耐NaClO脅迫,而且此現(xiàn)象在穩(wěn)定期更加明顯;當脅迫時間達到30 min時,突變株內(nèi)的活性氧水平增長率比野生株高出19.97%,進一步說明sigB基因?qū)aClO脅迫有一定的調(diào)控作用。RT-PCR結(jié)果表明,基因lmo1433、lmo2344和ohrR在野生株的表達水平顯著高于突變株(P0.01),而基因lmo0906、lmo2770的表達水平在野生株和缺失株中的差異并不明顯。結(jié)論:研究表明,單核細胞增生李斯特菌sigB基因?qū)aClO脅迫有一定的影響作用,且部分作用可能由激活部分調(diào)節(jié)機體氧化還原平衡的基因得以實現(xiàn)。
[Abstract]:Aim: to investigate the effect of sigB gene of Listeria monocytogenes on sodium hypochlorite (NaClO) stress. Methods: under the stress of NaClO(3 mmol / L, the tolerance of WaX12 and sigB knockout mutant and the level of reactive oxygen species in the wild strain were compared. The real-time quantitative polymerase chain reactionation (RT-PCR) technique was used to further observe the transcriptional level of lmo1433nlmo0906 / lmo234ohrRlmo2770 gene under stress time. Results: the phenotypic analysis of wild and sigB mutants showed that wild plants were more resistant to NaClO stress than the absent strains, and this phenomenon was more obvious in the stable period, and when the stress time reached 30 min, The growth rate of Ros in the mutant was 19.97 higher than that in the wild plant, which further indicated that sigB gene could regulate NaClO stress. RT-PCR results showed that the Ros level of the mutant was higher than that of the wild strain. The expression levels of lmo1433, lmo2344 and ohrR in wild plants were significantly higher than those in mutant strain P0.01, but the expression level of gene lmo0906 and lmo2770 was not significantly different between wild and absent plants. Conclusion: the results suggest that sigB gene of Listeria monocytogenes has a certain effect on NaClO stress, and part of the effect may be realized by activating genes that regulate the redox balance.
【作者單位】: 上海海洋大學食品學院農(nóng)業(yè)部水產(chǎn)品貯藏保鮮質(zhì)量安全風險評估實驗室水產(chǎn)品加工及貯藏工程技術(shù)研究中心;
【基金】:國家自然科學基金面上項目(31271870;31571917) 上海市科委計劃項目(14DZ1205100;14320502100);上海市科委工程中心建設項目(11DZ2280300) 上海市科技興農(nóng)重點攻關(guān)項目(滬農(nóng)科攻字2014第3-5號;2015第4-8號)
【分類號】:Q933
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