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利用三亲杂交的方法将带卡那抗性的Mini-Tn5转座子随机插入瓜类细菌性果斑病菌xjl12基因组DNA中,构建插入不同位点的突变体文库,通过信号分子高效检测菌株JZAI,筛选群体感应信号分子失活突变株.挑取2株野生株和筛选出的4株突变株,进行Southern杂交,结果表明,转座子Mini-Tn5以单拷贝随机诱变瓜类细菌性果斑病菌获得成功;对筛出的4株突变株进行了致病性和烟草过敏反应测试,发现突变株明显降低了致病性,且2株突变株(5-17,2-57)在烟草上无过敏反应.瓜类细菌性果斑病菌突变体文库的成功构建及筛选到不同突变位点的群体感应信号分子失活突变株,对从基因水平研究瓜类细菌性果斑病菌致病分子机理及群体感应相关基因奠定了基础. 相似文献
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Three new cyanobactins, trikoramides B (1)–D (3), have been isolated from the marine cyanobacterium, Symploca hydnoides, following a preliminary bioassay-guided isolation of the two most active polar fractions based on the brine shrimp toxicity assay. These new cyanobactins are new analogues of the previously reported cytotoxic trikoramide A (4) with differences mainly in the C-prenylated cyclotryptophan unit. Their planar structures were elucidated from their 1D and 2D NMR spectral data in combination with the HRMS/MS data. Marfey’s method, 2D NOESY NMR spectroscopic and ECD spectra analyses were used to determine the absolute stereochemistry of trikoramides B (1)–D (3). Trikoramides B (1) and D (3) exhibited cytotoxicity against MOLT-4 acute lymphoblastic leukemia cell line with IC50 values of 5.2 µM and 4.7 µM, respectively. Compounds 1 and 3 were also evaluated for their quorum-sensing inhibitory assay based on the Pseudomonas aeruginosa PAO1 lasB-gfp and rhlA-gfp bioreporter strains. Although trikoramide B (1) exhibited weak quorum-sensing inhibitory activity, the Br-containing trikoramide D (3) exhibited moderate to significant dose-dependent quorum-sensing inhibitory activities against PAO1 lasB-gpf and rhlA-gfp bioreporter strains with IC50 values of 19.6 µM and 7.3 µM, respectively. 相似文献
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石倩谢林香陈立志胡开辉张燎原 《广东农业科学》2014,41(10):77-80
微生物致病性在很大程度上受到群体感应系统的调控、筛选群体感应系统抑制剂被认为是解决微生物
致病性的有效方法之一。对17 种中草药进行了群体感应抑制剂的筛选、结果表明、其中8 种中草药能抑制
Chromobacterium violaceum CV026 产紫色色素的能力。试管梯度试验表明、这8 种中草药有明显的群体感应抑制效
果。浮游抑制试验结果表明、17 种中草药中的13 种可抑制铜绿假单胞菌的浮游。进一步对中草药中群体感应抑制剂
的分离鉴定、将为揭示细菌群体感应抑制剂的机制研究和应用奠定基础。 相似文献