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[Expression, purification and characterization of Rv3194c protein from Mycobacterium tuberculosis]. [结核分枝杆菌Rv3194c蛋白的表达、纯化和鉴定]。
Pub Date : 2016-12-04
Dongyue Zhao, Lili Lin, Fuli Wen

Objective: PDZ[Post-synaptic density-95 (PSD-95), Drosophilia tumor suppressor protein diskslarge-1 (DLG), the tight junction protein zonula occludentes 1 (ZO-1)] signal protein was encoded by Rv3194c gene from Mycobacterium tuberculosis, and its ability to adhere M. tuberculosis was studied.

Methods: Rv3194c protein was expressed in prokaryotic system. Rv3194c protein was separately incubated with hyaluronic acid, chondroitin sulfate and collagen Ι overnight at different temperature (37, 38, 39, 40℃). Then component changes of culture supernatant were tested by Western blot and ELISA.

Results: Western blot showed that Rv3194c protein expressed in prokaryotic system, with a molecular weight of about 35 kDa, was mainly in soluble form. Western blot showed that His-Rv3194c protein in supernatant of 39℃ experimental group was significantly less than that of other experimental groups (37, 38, 40℃)(***P<0.001). ELISA showed that hyaluronic acid, chondroitin sulfate and collagen Ι in supernatant of 39℃ experimental group was significantly less than that of other experimental groups (37, 38, 40℃)(***P<0.001).

Conclusion: For the first time it was affirmed that Rv3194c protein with detected activity of adhesions in this study will be targeted to the development of the new anti-M. tuberculosis drug.

目的:利用结核分枝杆菌Rv3194c基因编码PDZ[突触后密度-95 (PSD-95),果蝇肿瘤抑制蛋白diskslarge-1 (DLG),紧密连接蛋白occludentes 1 (ZO-1)]信号蛋白,并研究其对结核分枝杆菌的粘附能力。方法:在原核系统中表达Rv3194c蛋白。Rv3194c蛋白分别与透明质酸、硫酸软骨素和胶原蛋白Ι在不同温度(37、38、39、40℃)下孵育过夜。然后用Western blot和ELISA检测培养上清的成分变化。结果:Western blot结果显示,Rv3194c蛋白在原核系统中表达,分子量约为35 kDa,主要以可溶性形式表达。Western blot结果显示,39℃实验组上清中His-Rv3194c蛋白含量明显低于其他实验组(37、38、40℃)(*** p)。结论:本研究首次肯定了检测到黏附活性的Rv3194c蛋白将用于新型抗m抗体的开发。结核病药物。
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引用次数: 0
[Cloning, mutagenesis and symbiotic phenotype of three lipid transfer protein encoding genes from Mesorhizobium huakuii 7653R]. 3个脂质转移蛋白编码基因的克隆、诱变及共生表型研究[j]。
Pub Date : 2016-12-04
Yanan Li, Xiaobo Zeng, Xuejuan Zhou, Youguo Li

Objective: Lipid transfer protein superfamily is involved in lipid transport and metabolism. This study aimed to construct mutants of three lipid transfer protein encoding genes in Mesorhizobium huakuii 7653R, and to study the phenotypes and function of mutations during symbiosis with Astragalus sinicus.

Methods: We used bioinformatics to predict structure characteristics and biological functions of lipid transfer proteins, and conducted semi-quantitative and fluorescent quantitative real-time PCR to analyze the expression levels of target genes in free-living and symbiotic conditions. Using pK19mob insertion mutagenesis to construct mutants, we carried out pot plant experiments to observe symbiotic phenotypes.

Results: MCHK-5577, MCHK-2172 and MCHK-2779 genes encoding proteins belonged to START/RHO alpha_C/PITP/Bet_v1/CoxG/CalC (SRPBCC) superfamily, involved in lipid transport or metabolism, and were identical to M. loti at 95% level. Gene relative transcription level of the three genes all increased compared to free-living condition. We obtained three mutants. Compared with wild-type 7653R, above-ground biomass of plants and nodulenitrogenase activity induced by the three mutants significantly decreased.

Conclusion: Results indicated that lipid transfer protein encoding genes of Mesorhizobium huakuii 7653R may play important roles in symbiotic nitrogen fixation, and the mutations significantly affected the symbiotic phenotypes. The present work provided a basis to study further symbiotic function mechanism associated with lipid transfer proteins from rhizobia.

目的:脂质转运蛋白超家族参与脂质转运和代谢。本研究旨在构建华基中根瘤菌7653R中3个脂质转移蛋白编码基因的突变体,并研究突变体在与黄芪共生过程中的表型和功能。方法:利用生物信息学方法预测脂质转移蛋白的结构特征和生物学功能,并采用半定量和荧光定量实时PCR分析靶基因在自由生活和共生条件下的表达水平。利用pK19mob插入诱变技术构建突变体,进行盆栽实验,观察其共生表型。结果:MCHK-5577、MCHK-2172和MCHK-2779编码蛋白的基因属于START/RHO alpha_C/PITP/Bet_v1/CoxG/CalC (SRPBCC)超家族,参与脂质转运或代谢,与罗氏分枝杆菌在95%水平上相同。3个基因的相对转录水平均高于自由生活条件。我们得到了三个突变体。与野生型7653R相比,3个突变体诱导的植株地上生物量和根瘤氮素酶活性均显著降低。结论:华基中根菌7653R脂质转移蛋白编码基因可能在共生固氮过程中发挥重要作用,其突变显著影响共生表型。本研究为进一步研究根瘤菌脂质转移蛋白的共生功能机制提供了基础。
{"title":"[Cloning, mutagenesis and symbiotic phenotype of three lipid transfer protein encoding genes from Mesorhizobium huakuii 7653R].","authors":"Yanan Li,&nbsp;Xiaobo Zeng,&nbsp;Xuejuan Zhou,&nbsp;Youguo Li","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>Lipid transfer protein superfamily is involved in lipid transport and metabolism. This study aimed to construct mutants of three lipid transfer protein encoding genes in Mesorhizobium huakuii 7653R, and to study the phenotypes and function of mutations during symbiosis with Astragalus sinicus.</p><p><strong>Methods: </strong>We used bioinformatics to predict structure characteristics and biological functions of lipid transfer proteins, and conducted semi-quantitative and fluorescent quantitative real-time PCR to analyze the expression levels of target genes in free-living and symbiotic conditions. Using pK19mob insertion mutagenesis to construct mutants, we carried out pot plant experiments to observe symbiotic phenotypes.</p><p><strong>Results: </strong>MCHK-5577, MCHK-2172 and MCHK-2779 genes encoding proteins belonged to START/RHO alpha_C/PITP/Bet_v1/CoxG/CalC (SRPBCC) superfamily, involved in lipid transport or metabolism, and were identical to M. loti at 95% level. Gene relative transcription level of the three genes all increased compared to free-living condition. We obtained three mutants. Compared with wild-type 7653R, above-ground biomass of plants and nodulenitrogenase activity induced by the three mutants significantly decreased.</p><p><strong>Conclusion: </strong>Results indicated that lipid transfer protein encoding genes of Mesorhizobium huakuii 7653R may play important roles in symbiotic nitrogen fixation, and the mutations significantly affected the symbiotic phenotypes. The present work provided a basis to study further symbiotic function mechanism associated with lipid transfer proteins from rhizobia.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"56 12","pages":"1901-11"},"PeriodicalIF":0.0,"publicationDate":"2016-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36081984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Characterization of phenotype and expression regulation of an RND-type multidrug efflux pump in Mesorhizobium huakuii 7653R]. [一种rnd型多药外排泵在华奎中根菌7653R中的表型表征及表达调控]。
Pub Date : 2016-12-04
Ziling Liu, Jieli Peng, Youguo Li

Objective: To study the function of an RND family efflux pump encoded by MCHK_0866 and MCHK_0867 in Mesorhizobium huakuii 7653R.

Methods: Genetic organization of target genes was analyzed in genome. The change of growth was observed by measuring OD600. Drug sensitivity was detected by minimal inhibitory concentrations; relative transcription level of target genes was measured by RT-PCR. Transcript regulation of the efflux pump was validated by bacterial one-hybrid system.

Results: Proteins encoded by MCHK_0866 and MCHK_0867 formed an RND family efflux pump. The OD600 of growth curve reduced and it showed more sensitivity to nalidixic acid, tetracycline and SDS after disrupting the efflux pump. Genes relative transcription level increased in response to nalidixic acid treatment. Meanwhile, the downstream gene MCHK_0869 belongs to TetR family transcription factor and its expression product can interact with the promoter region of MCHK_0867.

Conclusion: The efflux pump is possibly associated with the transportation of nalidixic acid and affects rhizobial free-living growth. The pump is putatively regulated by a downstream local transcription factor.

目的:研究华葵中根菌7653R中MCHK_0866和MCHK_0867编码的RND家族外排泵的功能。方法:分析基因组中靶基因的遗传组织。通过测定OD600来观察生长变化。用最小抑菌浓度检测药物敏感性;RT-PCR检测靶基因的相对转录水平。通过细菌单杂交系统验证了外排泵的转录调控作用。结果:MCHK_0866和MCHK_0867编码的蛋白形成了RND家族外排泵。破坏外排泵后,生长曲线OD600降低,对萘啶酸、四环素和SDS的敏感性增强。钠地酸处理后基因相对转录水平升高。同时下游基因MCHK_0869属于TetR家族转录因子,其表达产物可与MCHK_0867的启动子区相互作用。结论:外排泵可能与钠酸的转运有关,影响根瘤菌的自由生长。据推测,这种泵是由下游的局部转录因子调节的。
{"title":"[Characterization of phenotype and expression regulation of an RND-type multidrug efflux pump in Mesorhizobium huakuii 7653R].","authors":"Ziling Liu,&nbsp;Jieli Peng,&nbsp;Youguo Li","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To study the function of an RND family efflux pump encoded by MCHK_0866 and MCHK_0867 in Mesorhizobium huakuii 7653R.</p><p><strong>Methods: </strong>Genetic organization of target genes was analyzed in genome. The change of growth was observed by measuring OD600. Drug sensitivity was detected by minimal inhibitory concentrations; relative transcription level of target genes was measured by RT-PCR. Transcript regulation of the efflux pump was validated by bacterial one-hybrid system.</p><p><strong>Results: </strong>Proteins encoded by MCHK_0866 and MCHK_0867 formed an RND family efflux pump. The OD600 of growth curve reduced and it showed more sensitivity to nalidixic acid, tetracycline and SDS after disrupting the efflux pump. Genes relative transcription level increased in response to nalidixic acid treatment. Meanwhile, the downstream gene MCHK_0869 belongs to TetR family transcription factor and its expression product can interact with the promoter region of MCHK_0867.</p><p><strong>Conclusion: </strong>The efflux pump is possibly associated with the transportation of nalidixic acid and affects rhizobial free-living growth. The pump is putatively regulated by a downstream local transcription factor.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"56 12","pages":"1876-82"},"PeriodicalIF":0.0,"publicationDate":"2016-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36083043","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Advances in CagA protein and CagAmediated pathogenesis of Helicobacter pylori-A review]. 【CagA蛋白及其介导的幽门螺杆菌发病机制研究进展综述】。
Pub Date : 2016-12-04
Xiukun Wan, Chunjie Liu

Helicobacter pylori (H. pylori) is a strong risk factor for gastric disease ranging from chronic gastritis to gastric cancer. But the mechanisms underlying the pathogenesis of H. pylori are still not completely understood.The cytotoxin-associated gene A (CagA) of H. pylori, an important virulence factor and the only bacterial oncoprotein, is extensively studied. CagA is delivered into gastric epithelial cells via type IV secretion of H. pylori. Upon delivery, CagA perturbs multiple host signaling pathways by interacting with the host signaling molecules, resulting in cytopathic effects and subsequent cell transformation. Some animal experiments also provide in vivo evidence for the oncogenic capacity of CagA. In this review, recent advances in the structural property, delivery manner and pathogenesis of CagA are summarized, which we hope could better explain the CagA-mediated pathogenesis of Helicobacter pylori and provide directions for the future approach.

幽门螺杆菌(Helicobacter pylori, H. pylori)是引起从慢性胃炎到胃癌等胃病的重要危险因素。但幽门螺杆菌的发病机制仍未完全了解。幽门螺杆菌细胞毒素相关基因A (CagA)是一种重要的毒力因子,也是唯一的细菌癌蛋白,目前已被广泛研究。CagA通过幽门螺杆菌IV型分泌物进入胃上皮细胞。在递送后,CagA通过与宿主信号分子相互作用干扰多种宿主信号通路,导致细胞病变效应和随后的细胞转化。一些动物实验也为CagA的致癌能力提供了体内证据。本文综述了近年来在CagA的结构特性、传递方式和发病机制等方面的研究进展,希望能更好地解释CagA介导的幽门螺杆菌发病机制,并为今后的研究提供方向。
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引用次数: 0
[Immunological characteristic comparison of different genetic polymorphism recombinant of FnBPA-A of bovine Staphylococcus aureus strain]. [牛金黄色葡萄球菌FnBPA-A不同基因多态性重组物的免疫学特性比较]。
Pub Date : 2016-12-04
Caidie Wang, Yan Su, Lingling Su, Haina Wei, Baojiang Zhang

Objective: To understand the impact of genetic polymorphism of FnBPA-A on the immune biological characteristics of Staphylococcus aureus.

Methods: Sequence of FnBPA-A of Staphylococcus aureus isolated from bovine in Xinjiang was analyzed and 8 different genetic polymorphism eukaryotic recombinants of FnBPA-A were constructed. C57BL/6 mice were immunized with these recombinant plasmids and mice sera were collected. Level of the immune protection of immunized mice was compared.

Results: GS801, GS819 and GS856 were on the same branch; GW10-1, GW20-2, GY288 and GY309 belong to the same branch; GY278 was on a different branch. For the challenge experiment, GW20-2, GS801, GS819, GS856 and GY288 showed better protection.

Conclusion: The genetic polymorphism of FnBPA-A could significantly affect the immune protection of immunized mice.

目的:了解FnBPA-A基因多态性对金黄色葡萄球菌免疫生物学特性的影响。方法:分析新疆牛金黄色葡萄球菌FnBPA-A基因序列,构建8个不同基因多态性的真核重组体。用重组质粒免疫C57BL/6小鼠,收集小鼠血清。比较免疫小鼠的免疫保护水平。结果:GS801、GS819和GS856在同一支上;GW10-1、GW20-2、GY288、GY309属于同一分支;GY278在不同的分支上。在攻毒实验中,GW20-2、GS801、GS819、GS856和GY288表现出较好的保护作用。结论:FnBPA-A基因多态性可显著影响免疫小鼠的免疫保护作用。
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引用次数: 0
[Characterization and microbial community shifts of rice strawdegrading microbial consortia]. [水稻秸秆降解菌群的特征及微生物群落迁移]。
Pub Date : 2016-12-04
Chunfang Wang, Shichun Ma, Yan Huang, Laiyan Liu, Hui Fan, Yu Deng

Objective: To study the relationship between microbial community and degradation rate of rice straw, we compared and analyzed cellulose-decomposing ability, microbial community structures and shifts of microbial consortia F1 and F2.

Methods: We determined exoglucanase activity by 3, 5-dinitrosalicylic acid colorimetry. We determined content of cellulose, hemicellulose and lignin in rice straw by Van Soest method, and calculated degradation rates of rice straw by the weight changes before and after a 10-day incubation. We analyzed and compared the microbial communities and functional microbiology shifts by clone libraries, Miseq analysis and real time-PCR based on the 16S rRNA gene and cel48 genes.

Results: Total degradation rate, cellulose, and hemicellulose degradation rate of microbial consortia F1 were significantly higher than that of F2. The variation trend of exoglucanase activity in both microbial consortia F1 and F2 was consistent with that of cel48 gene copies. Microbial diversity of F1 was complex with aerobic bacteria as dominant species, whereas that of F2 was simple with a high proportion of anaerobic cellulose decomposing bacteria in the later stage of incubation. In the first 4 days, unclassified Bacillales and Bacillus were dominant in both F1 and F2. The dominant species and abundance became different after 4-day incubation, Bacteroidetes and Firmicutes were dominant phyla of F1 and F2, respectively. Although Petrimonas and Pusillimonas were common dominant species in F1 and F2, abundance of Petrimonas in F2 (38.30%) was significantly higher than that in F1 (9.47%), and the abundance of Clostridiales OPB54 in F2 increased to 14.85% after 8-day incubation.

Conclusion: The abundance of cel48 gene related with cellulose degradation rate and exoglucanase activity, and cel48 gene has the potential as a molecular marker to monitor the process of cellulose degradation. Microbial community structure has a remarkable impact on the degradation efficiency of straw cellulose, and Petrimonas, Paenibacillus, Bacillales, Clostridiales were vital species for microbial consortia F1 and F2 decomposing rice straw.

目的:研究微生物群落与水稻秸秆降解率的关系,比较分析秸秆纤维素分解能力、微生物群落结构及群落F1和F2的变化。方法:采用3,5 -二硝基水杨酸比色法测定外葡聚糖酶活性。采用Van Soest法测定稻草中纤维素、半纤维素和木质素的含量,并通过培养10 d前后稻草重量的变化计算稻草的降解率。基于16S rRNA基因和cel48基因,采用克隆文库分析、Miseq分析和real - time-PCR分析比较了微生物群落和功能微生物变化。结果:微生物群落F1的总降解率、纤维素降解率和半纤维素降解率均显著高于F2。微生物群落F1和F2中外葡聚糖酶活性的变化趋势与cel48基因拷贝的变化趋势一致。F1的微生物多样性较为复杂,以好氧菌为优势菌种;F2的微生物多样性较为简单,培养后期厌氧纤维素分解菌比例较高。前4 d, F1和F2均以未分类芽孢杆菌和芽孢杆菌为主。孵育4 d后优势种和丰度有所不同,F1和F2的优势门分别为拟杆菌门和厚壁菌门。虽然在F1和F2中petronas和Pusillimonas是共同的优势种,但F2中petronas的丰度(38.30%)显著高于F1 (9.47%), F2中Clostridiales OPB54的丰度在培养8 d后增加到14.85%。结论:cel48基因丰度与纤维素降解速率和外葡聚糖酶活性有关,具有作为纤维素降解过程监测分子标记的潜力。微生物群落结构对秸秆纤维素的降解效率有显著影响,其中油单胞菌、Paenibacillus、芽孢杆菌、梭菌属(Clostridiales)是分解水稻秸秆的微生物群落F1和F2的重要物种。
{"title":"[Characterization and microbial community shifts of rice strawdegrading microbial consortia].","authors":"Chunfang Wang,&nbsp;Shichun Ma,&nbsp;Yan Huang,&nbsp;Laiyan Liu,&nbsp;Hui Fan,&nbsp;Yu Deng","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To study the relationship between microbial community and degradation rate of rice straw, we compared and analyzed cellulose-decomposing ability, microbial community structures and shifts of microbial consortia F1 and F2.</p><p><strong>Methods: </strong>We determined exoglucanase activity by 3, 5-dinitrosalicylic acid colorimetry. We determined content of cellulose, hemicellulose and lignin in rice straw by Van Soest method, and calculated degradation rates of rice straw by the weight changes before and after a 10-day incubation. We analyzed and compared the microbial communities and functional microbiology shifts by clone libraries, Miseq analysis and real time-PCR based on the 16S rRNA gene and cel48 genes.</p><p><strong>Results: </strong>Total degradation rate, cellulose, and hemicellulose degradation rate of microbial consortia F1 were significantly higher than that of F2. The variation trend of exoglucanase activity in both microbial consortia F1 and F2 was consistent with that of cel48 gene copies. Microbial diversity of F1 was complex with aerobic bacteria as dominant species, whereas that of F2 was simple with a high proportion of anaerobic cellulose decomposing bacteria in the later stage of incubation. In the first 4 days, unclassified Bacillales and Bacillus were dominant in both F1 and F2. The dominant species and abundance became different after 4-day incubation, Bacteroidetes and Firmicutes were dominant phyla of F1 and F2, respectively. Although Petrimonas and Pusillimonas were common dominant species in F1 and F2, abundance of Petrimonas in F2 (38.30%) was significantly higher than that in F1 (9.47%), and the abundance of Clostridiales OPB54 in F2 increased to 14.85% after 8-day incubation.</p><p><strong>Conclusion: </strong>The abundance of cel48 gene related with cellulose degradation rate and exoglucanase activity, and cel48 gene has the potential as a molecular marker to monitor the process of cellulose degradation. Microbial community structure has a remarkable impact on the degradation efficiency of straw cellulose, and Petrimonas, Paenibacillus, Bacillales, Clostridiales were vital species for microbial consortia F1 and F2 decomposing rice straw.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"56 12","pages":"1856-68"},"PeriodicalIF":0.0,"publicationDate":"2016-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36083041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Role of CRISPR/Cas systems in drugresistance and virulence and the effect of IS600 on the expression of cse2 in Shigella]. [CRISPR/Cas系统在志贺氏菌耐药和毒力中的作用以及IS600对cse2表达的影响]。
Pub Date : 2016-12-04
Lijuan Hong, Bing Zhang, Guangcai Duan, Wenjuan Liang, Yingfang Wang, Shuaiyin Chen, Haiyan Yang, Yuanlin Xi

Objective: To analyze the relationship between CRISPR/Cas system and drug-resistance, virulence. To investigate the effect of IS600 on the expression of CRISPR associated gene cse2 in Shigella.

Methods: CRISPR loci, CRISPR associated gene cse2, drug-resistant genes and virulent genes were detected by PCR in 33 Shigella strains; Trypan Blue counting test was used to detect bacterial virulence; Real-time PCR was used to detect relative mRNA expression of cse2; susceptibilities of Shigella strains were tested by agar diffusion method. Furthermore, we analyzed the relationship between CRISPR loci and drug-resistant genes, virulent genes. The effect of the IS600 on the expression of CRISPR associated gene cse2 was investigated.

Results: The mortality of Hela cells infected by Shigella with CRISPR1 loci was significantly lower (P<0.05) than those infected by Shigella without CRISPR1. The mRNA expression level of cse2 in group with IS600 was significantly (P<0.05) lower than that in group without IS600.

Conclusions: CRISPR loci were widely present in Shigella. Shigella without CRISPR1 has a higher pathogenicity. Due to the insertion of IS600, the mRNA expression level of cse2 was decreased in Shigella.

目的:分析CRISPR/Cas系统与耐药、毒力的关系。目的探讨IS600对志贺菌中CRISPR相关基因cse2表达的影响。方法:采用PCR检测33株志贺氏菌的CRISPR基因座、CRISPR相关基因cse2、耐药基因和毒力基因;台盼蓝计数法检测细菌毒力;Real-time PCR检测cse2 mRNA相对表达量;用琼脂扩散法检测志贺氏菌的药敏。进一步分析了CRISPR位点与耐药基因、毒力基因的关系。研究了IS600对CRISPR相关基因cse2表达的影响。结果:携带CRISPR1基因座的Hela细胞感染志贺氏菌后死亡率明显降低(p结论:CRISPR基因座在志贺氏菌中广泛存在。没有CRISPR1的志贺氏菌具有更高的致病性。由于IS600的插入,cse2 mRNA在志贺氏菌中的表达水平降低。
{"title":"[Role of CRISPR/Cas systems in drugresistance and virulence and the effect of IS600 on the expression of cse2 in Shigella].","authors":"Lijuan Hong,&nbsp;Bing Zhang,&nbsp;Guangcai Duan,&nbsp;Wenjuan Liang,&nbsp;Yingfang Wang,&nbsp;Shuaiyin Chen,&nbsp;Haiyan Yang,&nbsp;Yuanlin Xi","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To analyze the relationship between CRISPR/Cas system and drug-resistance, virulence. To investigate the effect of IS600 on the expression of CRISPR associated gene cse2 in Shigella.</p><p><strong>Methods: </strong>CRISPR loci, CRISPR associated gene cse2, drug-resistant genes and virulent genes were detected by PCR in 33 Shigella strains; Trypan Blue counting test was used to detect bacterial virulence; Real-time PCR was used to detect relative mRNA expression of cse2; susceptibilities of Shigella strains were tested by agar diffusion method. Furthermore, we analyzed the relationship between CRISPR loci and drug-resistant genes, virulent genes. The effect of the IS600 on the expression of CRISPR associated gene cse2 was investigated.</p><p><strong>Results: </strong>The mortality of Hela cells infected by Shigella with CRISPR1 loci was significantly lower (P<0.05) than those infected by Shigella without CRISPR1. The mRNA expression level of cse2 in group with IS600 was significantly (P<0.05) lower than that in group without IS600.</p><p><strong>Conclusions: </strong>CRISPR loci were widely present in Shigella. Shigella without CRISPR1 has a higher pathogenicity. Due to the insertion of IS600, the mRNA expression level of cse2 was decreased in Shigella.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"56 12","pages":"1912-23"},"PeriodicalIF":0.0,"publicationDate":"2016-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36081981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Polyphosphate and its physiological function in Mycobacteria - A review]. 多磷酸盐及其在分枝杆菌中的生理功能综述
Pub Date : 2016-12-04
Tingyu Shi, Xinggao Dong, Jianping Xie

Tuberculosis is still a global infectious disease. New drugs to shorten the course of treatment and new vaccines are the key points to control tuberculosis. The physiological study of Mycobacteria will contribute to the above-mentioned purposes. Polyphosphate plays an important role in the stress adaptation in bacteria. And there are two classes of enzymes:polyphosphate kinase and exopolyphosphatase involved in the polyphosphate metabolism to control the dynamic equilibrium of polyphosphate level in Mycobacteria. Present paper summarized the progress in metabolism and physiological roles of polyphosphate in Mycobacteria, to provide useful information for studying the physiological function of polyphosphate in Mycobacteria.

结核病仍然是一种全球性传染病。缩短疗程的新药和新疫苗是控制结核病的关键。分枝杆菌的生理学研究将有助于实现上述目的。多磷酸盐在细菌的逆境适应中起着重要的作用。多磷酸激酶和外多磷酸酶两类酶参与多磷酸代谢,控制分枝杆菌多磷酸水平的动态平衡。本文就多磷酸盐在分枝杆菌中的代谢及生理作用的研究进展进行综述,以期为研究多磷酸盐在分枝杆菌中的生理功能提供有益的信息。
{"title":"[Polyphosphate and its physiological function in Mycobacteria - A review].","authors":"Tingyu Shi,&nbsp;Xinggao Dong,&nbsp;Jianping Xie","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Tuberculosis is still a global infectious disease. New drugs to shorten the course of treatment and new vaccines are the key points to control tuberculosis. The physiological study of Mycobacteria will contribute to the above-mentioned purposes. Polyphosphate plays an important role in the stress adaptation in bacteria. And there are two classes of enzymes:polyphosphate kinase and exopolyphosphatase involved in the polyphosphate metabolism to control the dynamic equilibrium of polyphosphate level in Mycobacteria. Present paper summarized the progress in metabolism and physiological roles of polyphosphate in Mycobacteria, to provide useful information for studying the physiological function of polyphosphate in Mycobacteria.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"56 12","pages":"1840-6"},"PeriodicalIF":0.0,"publicationDate":"2016-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36083039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Identification and phytotoxic activity of fungus QTYC-51 from the gut of Pantala flavescens larvae]. [黄斑Pantala flavescens幼虫肠道真菌QTYC-51的鉴定及植物毒活性研究]。
Pub Date : 2016-12-04
Liping Jin, Yun Zhang, Xiao Xu, Xiaohui Xiao, Yinglao Zhang

Objective: To isolate the fungus with phytotoxic activity from the gut of Pantala flavescens larvae.

Methods: Strain QTYC-51 was identified by morphological observation and 5.8S rDNA-ITS sequence analysis. Petri dish bioassay was used to test the phytotoxic activity of fermentation broth and monomer compounds of strain QTYC-51 on Echinochloa crusgalli and Amaranthus retroflexus. Bioactive components were isolated from ethyl acetate extracts via chromatographic methods, and the structures were determined by mass spectrum and nuclear magnetic resonance analyses.

Results: QTYC-51 was identified as Paraconiothyrium sp.. The fermentation broth had good phytotoxic activity on radical growth of E. crusgalli and A. retroflexus with the inhibition rates of 76.9% and 56.5%, respectively. Five monomer compounds were purified from the fermentation products, including 1,8-dihydroxyanthraquinone, 1-hydroxy-10-methoxy-dibenz[b,e]oxepin-6,11-dione, hydroxyvertixanthone, globosuxanthone and 1,3,6,8-tetrahydroxyanthraquinone. At the concentration of 100 μg/mL, compound globosuxanthone was found to possess obvious phytotoxic effects on radical growth of E. crusgalli and A. retroflexus with the inhibition rates of 94.1% and 79.0%, respectively, which were comparable to that of positive control 2,4-dichlorophenoxyacetic acid. Compound 1-hydroxy-10-methoxy-dibenz[b,e] oxepin-6,11-dione showed potent phytotoxic activity against E. crusgalli and A. retroflexus with inhibition rates of 50.3% and 58.6%, respectively.

Conclusion: Strain QTYC-51 could be potentially developed as a microbial herbicide.

目的:从黄颡鱼幼虫肠道中分离出具有植物毒性活性的真菌。方法:采用形态学观察和5.8S rDNA-ITS序列分析对菌株QTYC-51进行鉴定。采用培养皿生物测定法测定菌株QTYC-51发酵液及其单体化合物对刺青藻(Echinochloa crusgalli)和苋菜(Amaranthus retroflexus)的毒活性。通过色谱分离得到乙酸乙酯提取物的活性成分,并通过质谱和核磁共振分析对其结构进行鉴定。结果:QTYC-51鉴定为甲状旁腺属。该发酵液对松果青霉和逆转录青霉的自由基生长具有良好的抑制作用,抑制率分别为76.9%和56.5%。从发酵产物中纯化出5个单体化合物,分别为1,8-二羟基蒽醌、1-羟基-10-甲氧基-二苯并[b,e]奥西平-6,11-二酮、羟基旋黄酮、球杉黄酮和1,3,6,8-四羟基蒽醌。结果表明,在100 μg/mL浓度下,复方球杉酮对地瓜和逆转录黄芪的自由基生长具有明显的抑制作用,抑制率分别为94.1%和79.0%,与阳性对照2,4-二氯苯氧乙酸相当。化合物1-羟基-10-甲氧基-二苯并[b,e]奥西平-6,11-二酮具有较强的植物毒活性,抑制率分别为50.3%和58.6%。结论:菌株QTYC-51具有开发微生物除草剂的潜力。
{"title":"[Identification and phytotoxic activity of fungus QTYC-51 from the gut of Pantala flavescens larvae].","authors":"Liping Jin,&nbsp;Yun Zhang,&nbsp;Xiao Xu,&nbsp;Xiaohui Xiao,&nbsp;Yinglao Zhang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To isolate the fungus with phytotoxic activity from the gut of Pantala flavescens larvae.</p><p><strong>Methods: </strong>Strain QTYC-51 was identified by morphological observation and 5.8S rDNA-ITS sequence analysis. Petri dish bioassay was used to test the phytotoxic activity of fermentation broth and monomer compounds of strain QTYC-51 on Echinochloa crusgalli and Amaranthus retroflexus. Bioactive components were isolated from ethyl acetate extracts via chromatographic methods, and the structures were determined by mass spectrum and nuclear magnetic resonance analyses.</p><p><strong>Results: </strong>QTYC-51 was identified as Paraconiothyrium sp.. The fermentation broth had good phytotoxic activity on radical growth of E. crusgalli and A. retroflexus with the inhibition rates of 76.9% and 56.5%, respectively. Five monomer compounds were purified from the fermentation products, including 1,8-dihydroxyanthraquinone, 1-hydroxy-10-methoxy-dibenz[b,e]oxepin-6,11-dione, hydroxyvertixanthone, globosuxanthone and 1,3,6,8-tetrahydroxyanthraquinone. At the concentration of 100 μg/mL, compound globosuxanthone was found to possess obvious phytotoxic effects on radical growth of E. crusgalli and A. retroflexus with the inhibition rates of 94.1% and 79.0%, respectively, which were comparable to that of positive control 2,4-dichlorophenoxyacetic acid. Compound 1-hydroxy-10-methoxy-dibenz[b,e] oxepin-6,11-dione showed potent phytotoxic activity against E. crusgalli and A. retroflexus with inhibition rates of 50.3% and 58.6%, respectively.</p><p><strong>Conclusion: </strong>Strain QTYC-51 could be potentially developed as a microbial herbicide.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"56 12","pages":"1869-75"},"PeriodicalIF":0.0,"publicationDate":"2016-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36083042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Construction of SPA7074-deficient mutant of biocontrol strain Streptomyces pactum Act12 and characterization of its secondary metabolites]. [生防菌株棕榈链霉菌Act12 spa7074缺失突变体的构建及其次生代谢产物的鉴定]。
Pub Date : 2016-12-04
Xuemei Duan, Feiyang Zhao, Xia Yan, Quanhong Xue, Xiaoxia Li, Bingjie Wen, Lianghui Jia, Hua Yan

Objective: To disrupt spa7074, which encodes a member of the TetR family transcriptional factors, in biocontrol strain Act12 and characterize the secondary metabolites in the mutant strain.

Methods: We disrupted the gene spa7074 by homologous recombination. The secondary metabolites of the mutant strain Δspa7074 and Act12 were detected by HPLC. The structure was analyzed by MS and NMR.

Results: Compared to the wild-type strain, the production of some unknown compounds in the mutant strain Δspa7074 increased obviously. We purified one of the compounds and identified as oligomycin D by MS and NMR analysis.

Conclusion: An oligomycin D-producing strain Δspa7074 was derived via genetic engineering.

目的:破坏生物防治菌株Act12中编码TetR家族转录因子的spa7074,并对其次生代谢产物进行表征。方法:采用同源重组法破坏spa7074基因。利用高效液相色谱法检测突变菌株Δspa7074和Act12的次生代谢产物。用质谱和核磁共振对其结构进行了分析。结果:与野生型菌株相比,突变株Δspa7074中一些未知化合物的产量明显增加。其中一个化合物经质谱和核磁共振鉴定为寡霉素D。结论:利用基因工程技术获得了一株产少霉素d的菌株Δspa7074。
{"title":"[Construction of SPA7074-deficient mutant of biocontrol strain Streptomyces pactum Act12 and characterization of its secondary metabolites].","authors":"Xuemei Duan,&nbsp;Feiyang Zhao,&nbsp;Xia Yan,&nbsp;Quanhong Xue,&nbsp;Xiaoxia Li,&nbsp;Bingjie Wen,&nbsp;Lianghui Jia,&nbsp;Hua Yan","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To disrupt spa7074, which encodes a member of the TetR family transcriptional factors, in biocontrol strain Act12 and characterize the secondary metabolites in the mutant strain.</p><p><strong>Methods: </strong>We disrupted the gene spa7074 by homologous recombination. The secondary metabolites of the mutant strain Δspa7074 and Act12 were detected by HPLC. The structure was analyzed by MS and NMR.</p><p><strong>Results: </strong>Compared to the wild-type strain, the production of some unknown compounds in the mutant strain Δspa7074 increased obviously. We purified one of the compounds and identified as oligomycin D by MS and NMR analysis.</p><p><strong>Conclusion: </strong>An oligomycin D-producing strain Δspa7074 was derived via genetic engineering.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":"56 12","pages":"1883-91"},"PeriodicalIF":0.0,"publicationDate":"2016-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36083044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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微生物学报
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