Archaea-An International Microbiological Journal最新文献

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Hydrogenotrophic Methanogenesis and Autotrophic Growth of Methanosarcina thermophila. 嗜热甲烷菌的亲氢产甲烷和自养生长。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2018-07-17 eCollection Date: 2018-01-01 DOI: 10.1155/2018/4712608
Nina Lackner, Anna Hintersonnleitner, Andreas Otto Wagner, Paul Illmer
{"title":"Hydrogenotrophic Methanogenesis and Autotrophic Growth of <i>Methanosarcina thermophila</i>.","authors":"Nina Lackner, Anna Hintersonnleitner, Andreas Otto Wagner, Paul Illmer","doi":"10.1155/2018/4712608","DOIUrl":"10.1155/2018/4712608","url":null,"abstract":"<p><p>Although Methanosarcinales are versatile concerning their methanogenic substrates, the ability of <i>Methanosarcina thermophila</i> to use carbon dioxide (CO<sub>2</sub>) for catabolic and anabolic metabolism was not proven until now. Here, we show that <i>M. thermophila</i> used CO<sub>2</sub> to perform hydrogenotrophic methanogenesis in the presence as well as in the absence of methanol. During incubation with hydrogen, the methanogen utilized the substrates methanol and CO<sub>2</sub> consecutively, resulting in a biphasic methane production. Growth exclusively from CO<sub>2</sub> occurred slowly but reproducibly with concomitant production of biomass, verified by DNA quantification. Besides verification through multiple transfers into fresh medium, the identity of the culture was confirmed by 16s RNA sequencing, and the incorporation of carbon atoms from <sup>13</sup>CO<sub>2</sub> into <sup>13</sup>CH<sub>4</sub> molecules was measured to validate the obtained data. New insights into the physiology of <i>M. thermophila</i> can serve as reference for genomic analyses to link genes with metabolic features in uncultured organisms.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2018-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6079545/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36409775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Characterization of Microbial Communities in Pilot-Scale Constructed Wetlands with Salicornia for Treatment of Marine Aquaculture Effluents. 用盐生植物处理海水养殖废水的试点规模建构湿地微生物群落特征。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2018-04-29 eCollection Date: 2018-01-01 DOI: 10.1155/2018/7819840
Xiaona Ma, Xingqiang Song, Xian Li, Songzhe Fu, Meng Li, Ying Liu
{"title":"Characterization of Microbial Communities in Pilot-Scale Constructed Wetlands with <i>Salicornia</i> for Treatment of Marine Aquaculture Effluents.","authors":"Xiaona Ma, Xingqiang Song, Xian Li, Songzhe Fu, Meng Li, Ying Liu","doi":"10.1155/2018/7819840","DOIUrl":"10.1155/2018/7819840","url":null,"abstract":"<p><p>Microorganisms play an essential role in the performance of constructed wetlands (CWs) for wastewater treatment. However, there has been limited discussion on the characteristics of microbial communities in CWs for treatment of effluents from marine recirculating aquaculture systems (RAS). This study is aimed at characterizing the microbial communities of pilot-scale CWs with <i>Salicornia bigelovii</i> for treatment of saline wastewater from a land-based Atlantic salmon RAS plant located in Northern China. Illumina high-throughput sequencing was employed to identify the profile of microbial communities of three CWs receiving wastewater under different total ammonia nitrogen (TAN) concentrations. Results of this study showed remarkable spatial variations in diversity and composition of microbial communities between roots and substrates in three CWs, with distinct response to different TAN concentrations. In particular, Proteobacteria, Firmicutes, Cyanobacteria, and Bacteroidetes were predominant in roots, while Cyanobacteria, Proteobacteria, Firmicutes, Verrucomicrobia, and Bacteroidetes were prevalent in substrates. Moreover, redundancy analysis indicated that specific functional genera, such as <i>Nitrosopumilus</i>, <i>Vibrio</i>, <i>Pseudoalteromonas</i>, <i>Nitrospina</i>, and <i>Planctomyces</i>, played key roles in the removal of nitrogen/phosphorus pollutants and growth of wetland plants. From a microorganism perspective, the findings of this study could contribute to better understanding of contaminants' removal mechanism and improved management of CWs for treatment of effluents from land-based marine aquaculture.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2018-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5949191/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36182024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Dynamics of Archaeal and Bacterial Communities in Response to Variations of Hydraulic Retention Time in an Integrated Anaerobic Fluidized-Bed Membrane Bioreactor Treating Benzothiazole Wastewater. 处理苯并噻唑废水的综合厌氧流化床膜生物反应器中古细菌和细菌群落随水力滞留时间变化的动态变化。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2018-04-29 eCollection Date: 2018-01-01 DOI: 10.1155/2018/9210534
Yue Li, Qi Hu, Da-Wen Gao
{"title":"Dynamics of Archaeal and Bacterial Communities in Response to Variations of Hydraulic Retention Time in an Integrated Anaerobic Fluidized-Bed Membrane Bioreactor Treating Benzothiazole Wastewater.","authors":"Yue Li, Qi Hu, Da-Wen Gao","doi":"10.1155/2018/9210534","DOIUrl":"10.1155/2018/9210534","url":null,"abstract":"<p><p>An integrated anaerobic fluidized-bed membrane bioreactor (IAFMBR) was investigated to treat synthetic high-strength benzothiazole wastewater (50 mg/L) at a hydraulic retention time (HRT) of 24, 18, and 12 h. The chemical oxygen demand (COD) removal efficiency (from 93.6% to 90.9%), the methane percentage (from 70.9% to 69.27%), and the methane yield (from 0.309 m<sup>3</sup> CH<sub>4</sub>/kg·COD<sub>removed</sub> to 0.316 m<sup>3</sup> CH<sub>4</sub>/kg·COD<sub>removed</sub>) were not affected by decreasing HRTs. However, it had an adverse effect on membrane fouling (decreasing service period from 5.3 d to 3.2 d) and benzothiazole removal efficiency (reducing it from 97.5% to 82.3%). Three sludge samples that were collected on day 185, day 240, and day 297 were analyzed using an Illumina® MiSeq platform. It is striking that the dominant genus of archaea was always <i>Methanosaeta</i> despite of HRTs. The proportions of <i>Methanosaeta</i> were 80.6% (HRT 24), 91.9% (HRT 18), and 91.2% (HRT 12). The dominant bacterial genera were <i>Clostridium</i> in proportions of 23.9% (HRT 24), 16.4% (HRT 18), and 15.3% (HRT 12), respectively.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2018-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5949192/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36181097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Inorganic Polyphosphate, Exopolyphosphatase, and Pho84-Like Transporters May Be Involved in Copper Resistance in Metallosphaera sedula DSM 5348T. 无机多磷酸盐、外多磷酸酶和pho84样转运蛋白可能参与了seula Metallosphaera DSM 5348T的铜抗性。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2018-03-05 eCollection Date: 2018-01-01 DOI: 10.1155/2018/5251061
Matías Rivero, Constanza Torres-Paris, Rodrigo Muñoz, Ricardo Cabrera, Claudio A Navarro, Carlos A Jerez
{"title":"Inorganic Polyphosphate, Exopolyphosphatase, and <i>Pho84</i>-Like Transporters May Be Involved in Copper Resistance in <i>Metallosphaera sedula</i> DSM 5348<sup>T</sup>.","authors":"Matías Rivero,&nbsp;Constanza Torres-Paris,&nbsp;Rodrigo Muñoz,&nbsp;Ricardo Cabrera,&nbsp;Claudio A Navarro,&nbsp;Carlos A Jerez","doi":"10.1155/2018/5251061","DOIUrl":"https://doi.org/10.1155/2018/5251061","url":null,"abstract":"<p><p>Polyphosphates (PolyP) are linear polymers of orthophosphate residues that have been proposed to participate in metal resistance in bacteria and archaea. In addition of having a CopA/CopB copper efflux system, the thermoacidophilic archaeon <i>Metallosphaera sedula</i> contains electron-dense PolyP-like granules and a putative exopolyphosphatase (PPX <i><sub>Msed</sub></i> , <i>Msed_0891</i>) and four presumed <i>pho84</i>-like phosphate transporters (<i>Msed_0846</i>, <i>Msed_0866</i>, <i>Msed_1094</i>, and <i>Msed_1512</i>) encoded in its genome. In the present report, the existence of a possible PolyP-based copper-resistance mechanism in <i>M. sedula</i> DSM 5348<sup>T</sup> was evaluated. <i>M. sedula</i> DSM 5348<sup>T</sup> accumulated high levels of phosphorous in the form of granules, and its growth was affected in the presence of 16 mM copper. PolyP levels were highly reduced after the archaeon was subjected to an 8 mM CuSO<sub>4</sub> shift. PPX <i><sub>Msed</sub></i> was purified, and the enzyme was found to hydrolyze PolyP <i>in vitro</i>. Essential residues for catalysis of PPX <i><sub>Msed</sub></i> were E111 and E113 as shown by a site-directed mutagenesis of the implied residues. Furthermore, <i>M. sedula ppx</i>, <i>pho84</i>-like, and <i>copTMA</i> genes were upregulated upon copper exposure, as determined by qRT-PCR analysis. The results obtained support the existence of a PolyP-dependent copper-resistance system that may be of great importance in the adaptation of this thermoacidophilic archaeon to its harsh environment.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2018-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2018/5251061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36041406","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 13
Community and Proteomic Analysis of Anaerobic Consortia Converting Tetramethylammonium to Methane. 四甲基铵转化为甲烷的厌氧菌群落及蛋白质组学分析。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2017-12-17 eCollection Date: 2017-01-01 DOI: 10.1155/2017/2170535
Wei-Yu Chen, Lucia Kraková, Jer-Horng Wu, Domenico Pangallo, Lenka Jeszeová, Bing Liu, Hidenari Yasui
{"title":"Community and Proteomic Analysis of Anaerobic Consortia Converting Tetramethylammonium to Methane.","authors":"Wei-Yu Chen,&nbsp;Lucia Kraková,&nbsp;Jer-Horng Wu,&nbsp;Domenico Pangallo,&nbsp;Lenka Jeszeová,&nbsp;Bing Liu,&nbsp;Hidenari Yasui","doi":"10.1155/2017/2170535","DOIUrl":"https://doi.org/10.1155/2017/2170535","url":null,"abstract":"<p><p>Tetramethylammonium-degrading methanogenic consortia from a complete-mixing suspended sludge (CMSS) and an upflow anaerobic sludge blanket (UASB) reactors were studied using multiple PCR-based molecular techniques and shotgun proteomic approach. The prokaryotic 16S rRNA genes of the consortia were analyzed by quantitative PCR, high-throughput sequencing, and DGGE-cloning methods. The results showed that methanogenic <i>archaea</i> were highly predominant in both reactors but differed markedly according to community structure. Community and proteomic analysis revealed that <i>Methanomethylovorans</i> and <i>Methanosarcina</i> were the major players for the demethylation of methylated substrates and methane formation through the reduction pathway of methyl-S-CoM and possibly, acetyl-CoA synthase/decarbonylase-related pathways. Unlike high dominance of one <i>Methanomethylovorans</i> population in the CMSS reactor, diverse methylotrophic <i>Methanosarcina</i> species inhabited in syntrophy-like association with hydrogenotrophic <i>Methanobacterium</i> in the granular sludge of UASB reactor. The overall findings indicated the reactor-dependent community structures of quaternary amines degradation and provided microbial insight for the improved understanding of engineering application.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2017-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/2170535","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35785738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 5
A Flexible System for Cultivation of Methanococcus and Other Formate-Utilizing Methanogens. 一种灵活的产甲烷球菌及其他利用甲酸的产甲烷菌培养系统。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2017-11-19 eCollection Date: 2017-01-01 DOI: 10.1155/2017/7046026
Feng Long, Liangliang Wang, Boguslaw Lupa, William B Whitman
{"title":"A Flexible System for Cultivation of <i>Methanococcus</i> and Other Formate-Utilizing Methanogens.","authors":"Feng Long,&nbsp;Liangliang Wang,&nbsp;Boguslaw Lupa,&nbsp;William B Whitman","doi":"10.1155/2017/7046026","DOIUrl":"https://doi.org/10.1155/2017/7046026","url":null,"abstract":"<p><p>Many hydrogenotrophic methanogens use either H<sub>2</sub> or formate as the major electron donor to reduce CO<sub>2</sub> for methane production. The conventional cultivation of these organisms uses H<sub>2</sub> and CO<sub>2</sub> as the substrate with frequent replenishment of gas during growth. H<sub>2</sub> is explosive and requires an expensive gassing system to handle safely. Formate is as an ideal alternative substrate from the standpoints of both economy and safety but leads to large changes in the culture pH during growth. Here, we report that glycylglycine is an inexpensive and nontoxic buffer suitable for growth of <i>Methanococcus maripaludis</i> and <i>Methanothermococcus okinawensis</i>. This cultivation system is suitable for growth on liquid as well as solid medium in serum bottles. Moreover, it allows cultivation of liter scale cultures without expensive fermentation equipment. This formate cultivation system provides an inexpensive and flexible alternative for the growth of formate-utilizing, hydrogenotrophic methanogens.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2017-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/7046026","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35749190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 27
Expression, Purification, and Characterization of (R)-Sulfolactate Dehydrogenase (ComC) from the Rumen Methanogen Methanobrevibacter millerae SM9. 瘤胃产甲烷菌(R)-硫代乳酸脱氢酶(ComC)的表达、纯化和鉴定
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2017-11-06 eCollection Date: 2017-01-01 DOI: 10.1155/2017/5793620
Yanli Zhang, Linley R Schofield, Carrie Sang, Debjit Dey, Ron S Ronimus
{"title":"Expression, Purification, and Characterization of (<i>R</i>)-Sulfolactate Dehydrogenase (ComC) from the Rumen Methanogen <i>Methanobrevibacter millerae</i> SM9.","authors":"Yanli Zhang,&nbsp;Linley R Schofield,&nbsp;Carrie Sang,&nbsp;Debjit Dey,&nbsp;Ron S Ronimus","doi":"10.1155/2017/5793620","DOIUrl":"https://doi.org/10.1155/2017/5793620","url":null,"abstract":"<p><p>(<i>R</i>)-Sulfolactate dehydrogenase (EC 1.1.1.337), termed ComC, is a member of an NADH/NADPH-dependent oxidoreductase family of enzymes that catalyze the interconversion of 2-hydroxyacids into their corresponding 2-oxoacids. The ComC reaction is reversible and in the biosynthetic direction causes the conversion of (<i>R</i>)-sulfolactate to sulfopyruvate in the production of coenzyme M (2-mercaptoethanesulfonic acid). Coenzyme M is an essential cofactor required for the production of methane by the methyl-coenzyme M reductase complex. ComC catalyzes the third step in the first established biosynthetic pathway of coenzyme M and is also involved in methanopterin biosynthesis. In this study, ComC from <i>Methanobrevibacter millerae</i> SM9 was cloned and expressed in <i>Escherichia coli</i> and biochemically characterized. Sulfopyruvate was the preferred substrate using the reduction reaction, with 31% activity seen for oxaloacetate and 0.2% seen for <i>α</i>-ketoglutarate. Optimal activity was observed at pH 6.5. The apparent <i>K</i><sub>M</sub> for coenzyme (NADH) was 55.1 <i>μ</i>M, and for sulfopyruvate, it was 196 <i>μ</i>M (for sulfopyruvate the <i>V</i><sub>max</sub> was 93.9 <i>μ</i>mol min<sup>-1</sup> mg<sup>-1</sup> and <i>k</i><sub>cat</sub> was 62.8 s<sup>-1</sup>). The critical role of ComC in two separate cofactor pathways makes this enzyme a potential means of developing methanogen-specific inhibitors for controlling ruminant methane emissions which are increasingly being recognized as contributing to climate change.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2017-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/5793620","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35650604","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 5
Growth Characteristics of Methanomassiliicoccus luminyensis and Expression of Methyltransferase Encoding Genes. 鲁米尼氏甲烷菌(Methanomassiliicoccus luminyensis)的生长特性和甲基转移酶编码基因的表达。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2017-11-02 eCollection Date: 2017-01-01 DOI: 10.1155/2017/2756573
Lena Kröninger, Jacqueline Gottschling, Uwe Deppenmeier
{"title":"Growth Characteristics of <i>Methanomassiliicoccus luminyensis</i> and Expression of Methyltransferase Encoding Genes.","authors":"Lena Kröninger, Jacqueline Gottschling, Uwe Deppenmeier","doi":"10.1155/2017/2756573","DOIUrl":"10.1155/2017/2756573","url":null,"abstract":"<p><p>DNA sequence analysis of the human gut revealed the presence a seventh order of methanogens referred to as Methanomassiliicoccales. <i>Methanomassiliicoccus luminyensis</i> is the only member of this order that grows in pure culture. Here, we show that the organism has a doubling time of 1.8 d with methanol + H<sub>2</sub> and a growth yield of 2.4 g dry weight/mol CH<sub>4</sub>. <i>M. luminyensis</i> also uses methylamines + H<sub>2</sub> (monomethylamine, dimethylamine, and trimethylamine) with doubling times of 2.1-2.3 d. Similar cell yields were obtained with equimolar concentrations of methanol and methylamines with respect to their methyl group contents. The transcript levels of genes encoding proteins involved in substrate utilization indicated increased amounts of mRNA from the <i>mtaBC2</i> gene cluster in methanol-grown cells. When methylamines were used as substrates, mRNA of the <i>mtb/mtt</i> operon and of the <i>mtmBC1</i> cluster were found in high abundance. The transcript level of <i>mtaC2</i> was almost identical in methanol- and methylamine-grown cells, indicating that genes for methanol utilization were constitutively expressed in high amounts. The same observation was made with resting cells where methanol always yielded the highest CH<sub>4</sub> production rate independently from the growth substrate. Hence, <i>M. luminyensis</i> is adapted to habitats that provide methanol + H<sub>2</sub> as substrates.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2017-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688252/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35332952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Development of the Multiple Gene Knockout System with One-Step PCR in Thermoacidophilic Crenarchaeon Sulfolobus acidocaldarius. 嗜热嗜酸绿藻(Crenarchaeon Sulfolobus acidocalarius)一步PCR多基因敲除系统的建立
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2017-10-31 eCollection Date: 2017-01-01 DOI: 10.1155/2017/7459310
Shoji Suzuki, Norio Kurosawa
{"title":"Development of the Multiple Gene Knockout System with One-Step PCR in Thermoacidophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i>.","authors":"Shoji Suzuki,&nbsp;Norio Kurosawa","doi":"10.1155/2017/7459310","DOIUrl":"https://doi.org/10.1155/2017/7459310","url":null,"abstract":"<p><p>Multiple gene knockout systems developed in the thermoacidophilic crenarchaeon <i>Sulfolobus acidocaldarius</i> are powerful genetic tools. However, plasmid construction typically requires several steps. Alternatively, PCR tailing for high-throughput gene disruption was also developed in <i>S. acidocaldarius</i>, but repeated gene knockout based on PCR tailing has been limited due to lack of a genetic marker system. In this study, we demonstrated efficient homologous recombination frequency (2.8 × 10<sup>4</sup> ± 6.9 × 10<sup>3</sup> colonies/<i>μ</i>g DNA) by optimizing the transformation conditions. This optimized protocol allowed to develop reliable gene knockout via double crossover using short homologous arms and to establish the multiple gene knockout system with one-step PCR (MONSTER). In the MONSTER, a multiple gene knockout cassette was simply and rapidly constructed by one-step PCR without plasmid construction, and the PCR product can be immediately used for target gene deletion. As an example of the applications of this strategy, we successfully made a DNA photolyase- (<i>phr-</i>) and arginine decarboxylase- (<i>argD-</i>) deficient strain of <i>S. acidocaldarius</i>. In addition, an agmatine selection system consisting of an agmatine-auxotrophic strain and <i>argD</i> marker was also established. The MONSTER provides an alternative strategy that enables the very simple construction of multiple gene knockout cassettes for genetic studies in <i>S. acidocaldarius</i>.</p>","PeriodicalId":49105,"journal":{"name":"Archaea-An International Microbiological Journal","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2017-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/7459310","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35241247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 13
Archaeal Diversity and CO2 Fixers in Carbonate-/Siliciclastic-Rock Groundwater Ecosystems. 碳酸盐/硅塑料-岩石地下水生态系统中古细菌多样性和CO2固定物。
IF 2.4 4区 生物学
Archaea-An International Microbiological Journal Pub Date : 2017-06-13 eCollection Date: 2017-01-01 DOI: 10.1155/2017/2136287
Cassandre Sara Lazar, Wenke Stoll, Robert Lehmann, Martina Herrmann, Valérie F Schwab, Denise M Akob, Ali Nawaz, Tesfaye Wubet, François Buscot, Kai-Uwe Totsche, Kirsten Küsel
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引用次数: 27
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