<meta charset="utf8"></head><body>与真菌伴侣共培养蔗糖分泌蓝细菌的方案

<table><tbody><tr><td>10x Ph1 objective</td><td>Carl Zeiss AG</td><td>46 04 01-9904</td><td></td></tr><tr><td>525/40 nm Bandpass Filter</td><td>Beckman Coulter, Inc.</td><td>A01-1-0051</td><td>FITC filter (in 488 nm laser)</td></tr><tr><td>660/10 nm Bandpass Filter</td><td>Beckman Coulter, Inc.</td><td>A01-1-0055</td><td>APC channel (in 638 nm laser)</td></tr><tr><td>710/50 nm Bandpass Filter</td><td>Beckman Coulter, Inc.</td><td>B71092</td><td>PC5.5 channel (in 561 nm laser)</td></tr><tr><td>Accuvettes for Multisizer 4e</td><td>Beckman Coulter, Inc.</td><td>A35473</td><td>Sample cups with lids</td></tr><tr><td>Aperture tube 30 &amp;mu;M for Multisizer 4e</td><td>Beckman Coulter, Inc.</td><td>C92760</td><td>Capillary</td></tr><tr><td>Autoclave</td><td>Lab Associates</td><td>EQM-ARBP</td><td>for sterilisation of material</td></tr><tr><td>Baffled flasks ROTILABO&amp;reg;, Straight neck, 250 ml</td><td>Carl Roth GmbH + Co. KG</td><td>LY95.1</td><td></td></tr><tr><td>BG11 / CoYBG11 components</td><td>different suppliers</td><td></td><td>see Table 1&amp;nbsp;</td></tr><tr><td>Bottle top sterile filter</td><td>Sigmaaldrich</td><td>Z222593</td><td>to sterile media if you dont want to use autoclave</td></tr><tr><td>Capillary 45 &amp;micro;m</td><td>Omni Life Science</td><td>OLS 5651738</td><td></td></tr><tr><td>CASY Cell Counter Model TTC</td><td>Schaerfe Systems</td><td></td><td></td></tr><tr><td>CASY cups</td><td>Omni Life Science</td><td>OLS 5651794</td><td></td></tr><tr><td>CASYclean</td><td>Omni Life Science</td><td>OLS 5651786</td><td>Cleaning solution</td></tr><tr><td>CASYton solution</td><td>Omni Life Science</td><td>OLS 5651808</td><td>Isotonic solution&amp;nbsp;</td></tr><tr><td>Centrifuge tubes (50 mL)</td><td>Sigmaaldrich</td><td>CLS430828</td><td></td></tr><tr><td>CO&lt;sub&gt;2&lt;/sub&gt;</td><td>BOC</td><td>270182-L</td><td>to supply cyanobacteria axenic cultures and co-cultures</td></tr><tr><td>CONTRAD&amp;reg; 70</td><td>Beckman Coulter, Inc.</td><td>81911</td><td>Deep clean solution</td></tr><tr><td>Cover glass 20 &amp;times; 26 x 0.4 mm</td><td>VWR International GmbH</td><td>631-1190</td><td>Cover glass for counting chamber</td></tr><tr><td>Cuvettes</td><td>Sarstedt&amp;nbsp;</td><td>67.742</td><td></td></tr><tr><td>Cyanobacteria BG-11 Freshwater Solution</td><td>Merck KGaA</td><td>C3061</td><td></td></tr><tr><td>CytExpert Software v. 2.6</td><td>Beckman Coulter, Inc.</td><td></td><td>Cytometer application software</td></tr><tr><td>CytoFLEX Sheath Fluid</td><td>Beckman Coulter, Inc.</td><td>B51503</td><td>Sheat fluid</td></tr><tr><td>Cytometer CytoFLEX S</td><td>Beckman Coulter, Inc.</td><td>BE51180</td><td></td></tr><tr><td>DxH Cleaner for Multisizer 4e</td><td>Beckman Coulter, Inc.</td><td>628022</td><td>Cleaning solution</td></tr><tr><td>Eppendorf tube centrifuge (small)</td><td>Labstac</td><td>CEN16-15</td><td>to centrifuge less than 2mL of culture</td></tr><tr><td>Ethanol</td><td>Carl Roth GmbH + Co. KG</td><td>K928.1</td><td></td></tr><tr><td>FACS tubes</td><td>Beckman Coulter, Inc.</td><td>2523749</td><td>tubes for semi-automatic mode</td></tr><tr><td>FlowClean Cleaning Agent, 500ml</td><td>Beckman Coulter, Inc.</td><td>C48093</td><td>Daily clean solution</td></tr><tr><td>Graduated cylinders</td><td>Sigmaaldrich</td><td>Z131121</td><td>for preparation of media</td></tr><tr><td>HEPPSO</td><td>Merck KGaA</td><td>R426725</td><td></td></tr><tr><td>Infors MultiTron photoincubator</td><td>Infors AG</td><td></td><td>white LED light</td></tr><tr><td>IPTG</td><td>Merck KGaA</td><td>I6758</td><td>Prepare aliquots, store at -20&amp;deg;C, thaw and keep on ice&amp;nbsp;</td></tr><tr><td>Isoton II for Multisizer 4e</td><td>Beckman Coulter, Inc.</td><td>8448011</td><td>Isotonic solution&amp;nbsp;</td></tr><tr><td>Kaluza v. 2.2</td><td>Beckman Coulter, Inc.</td><td>B16406</td><td>Cytometry data analysis software</td></tr><tr><td>KimWipes</td><td>VWR International GmbH</td><td>115-2221</td><td>Lint-free tissues</td></tr><tr><td>KOH</td><td>Merck KGaA</td><td>221473</td><td></td></tr><tr><td>KPO&lt;sub&gt;3&lt;/sub&gt;</td><td>Noah Chemicals</td><td>7790-53-6</td><td></td></tr><tr><td>Measuring beakers</td><td>VWR</td><td>213-3747</td><td></td></tr><tr><td>Micro test plate, 96 well, slip-on lid, flat base, PS, transparent</td><td>SARSTEDT AG &amp;amp; Co. KG</td><td>82.1581.001</td><td></td></tr><tr><td>Microscope base</td><td>Carl Zeiss AG</td><td>47 09 18-9902/16</td><td></td></tr><tr><td>Microsope head</td><td>Carl Zeiss AG</td><td>47 30 14</td><td></td></tr><tr><td>Milli-Q water purification system</td><td>Merck Millipore</td><td>C85358</td><td></td></tr><tr><td>Mulitisizer 4e particle counter</td><td>Beckman Coulter, Inc.</td><td>B23005</td><td>Alternative to the CASY Cell Counter</td></tr><tr><td>Neubauer improved, depth 0.02 mm</td><td>Assistent Germany</td><td></td><td>Counting chamber</td></tr><tr><td>Neubauer improved, depth 0.1 mm</td><td>Paul Marienfeld GmbH &amp;amp; Co. KG</td><td>640010</td><td>Counting chamber</td></tr><tr><td>Ready to Use Daily QC Fluorospheres</td><td>Beckman Coulter, Inc.</td><td>C65719</td><td>Reference for QC</td></tr><tr><td>SCHOTT bottles</td><td>Dutscher</td><td>90347</td><td>for storage</td></tr><tr><td>Specord&amp;reg;200 Plus Spectrophotometer</td><td>Analytic Jena GmbH</td><td></td><td>OD600/750</td></tr><tr><td>Sucrose</td><td>Merck KGaA</td><td>84100</td><td></td></tr><tr><td>Table top centrifuge (big)</td><td>Labstac</td><td>CEN18-06R</td><td>collect culture biomass</td></tr><tr><td>Yeast Nitrogen Base, without amino acids, without ammonium sulfate</td><td>Thermo Fisher Scientific Inc.</td><td>11743014</td><td></td></tr></tbody></table>

assembly and quantification of co-cultures combining heterotrophic yeast with phototrophic sugar-secreting cyanobacteria

<meta charset="utf8"></head><body>异养酵母与光养分泌糖的蓝细菌相结合的共培养物的组装和定量

异养酵母与光养分泌糖的蓝细菌相结合的共培养物的组装和定量

With the increasing demand for sustainable biotechnologies, mixed consortia containing a phototrophic microbe and heterotrophic partner species are being ...

assembly-quantification-co-cultures-combining-heterotrophic-yeast

Nanyang Technological University

Research

JoVE Journal

Bioengineering

Assembly and Quantification of Co-Cultures Combining Heterotrophic Yeast with Phototrophic Sugar-Secreting Cyanobacteria

662 次观看.  Heinrich Heine University Düsseldorf. 该协议为共培养物的设置和定量监测提供了全面的指南，包括光合自养分泌糖的蓝细菌和异养酵母。 

视频: 异养酵母与光养分泌糖的蓝细菌相结合的共培养物的组装和定量

Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry

Carbon-based nanomaterials, like carbon nanotubes (CNTs), belong to this type of nanoparticles which are very difficult to discriminate from carbon-rich cell structures and de facto there is still no quantitative method to assess their distribution at cell and tissue levels. What we propose here is an innovative method allowing the detection and quantification of CNTs in cells using a multispectral imaging flow cytometer (ImageStream, Amnis). This newly developed device integrates both a high-throughput of cells and high resolution imaging, providing thus images for each cell directly in flow and therefore statistically relevant image analysis. Each cell image is acquired on bright-field (BF), dark-field (DF), and fluorescent channels, giving access respectively to the level and the distribution of light absorption, light scattered and fluorescence for each cell. The analysis consists then in a pixel-by-pixel comparison of each image, of the 7,000-10,000 cells acquired for each condition of the experiment. Localization and quantification of CNTs is made possible thanks to some particular intrinsic properties of CNTs: strong light absorbance and scattering; indeed CNTs appear as strongly absorbed dark spots on BF and bright spots on DF with a precise colocalization.
This methodology could have a considerable impact on studies about interactions between nanomaterials and cells given that this protocol is applicable for a large range of nanomaterials, insofar as they are capable of absorbing (and/or scattering) strongly enough the light.

In this study, the main purpose was to monitor the cellular uptake and eventual exocytosis of carbon nanotubes (CNTs). From this perspective, we proposed here a unique method using multispectral imaging flow cytometry allowing a quantification and localization of CNTs in a statistically relevant number of cells.

Carbon-based nanomaterials, like carbon nanotubes (CNTs), belong to this type of nanoparticles which are very difficult to discriminate from carbon-rich ...

科研

生物工程

Cultivation of Green Microalgae in Bubble Column Photobioreactors and an Assay for Neutral Lipids

There is significant interest in the study of microalgae for engineering applications such as the production of biofuels, high value products, and for the treatment of wastes. As most new research efforts begin at laboratory scale, there is a need for cost-effective methods for culturing microalgae in a reproducible manner. Here, we communicate an effective approach to culture microalgae in laboratory-scale photobioreactors, and to measure the growth and neutral lipid content of that algae. Instructions are also included on how to set up the photobioreactor system. Although the example organisms are species of Chlorella and Auxenochlorella, this system can be adapted to cultivate a wide range of microalgae, including co-cultures of algae with non-algae species. Stock cultures are first grown in bottles to produce inoculum for the photobioreactor system. Algae inoculum is concentrated and transferred to photobioreactors for cultivation in batch mode. Samples are collected daily for the optical density readings. At the end of the batch culture, cells are harvested by centrifuge, washed, and freeze dried to obtain a final dry weight concentration. The final dry weight concentration is used to create a correlation between the optical density and the dry weight concentration. A modified Folch method is subsequently used to extract total lipids from the freeze-dried biomass and the extract is assayed for its neutral lipid content using a microplate assay. This assay has been published previously but protocol steps were included here to highlight critical steps in the procedure where errors frequently occur. The bioreactor system described here fills a niche between simple flask cultivation and fully-controlled commercial bioreactors. Even with only 3-4 biological replicates per treatment, our approach to culturing algae leads to tight standard deviations in the growth and lipid assays.

Here, we present a protocol to construct lab-scale bubble column photobioreactors and use them to culture microalgae. It also provides a method for the determination of the culture growth rate and neutral lipid content.

气泡柱光生物反应器中绿色微藻的培养及中性脂质的测定

There is significant interest in the study of microalgae for engineering applications such as the production of biofuels, high value products, and for the ...

环境科学

ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly

The extracellular matrix (ECM) in tissues is synthesized and assembled by cells to form a 3D fibrillar, protein network with tightly regulated fiber diameter, composition and organization. In addition to providing structural support, the physical and chemical properties of the ECM play an important role in multiple cellular processes including adhesion, differentiation, and apoptosis. In vivo, the ECM is assembled by exposing cryptic self-assembly (fibrillogenesis) sites within proteins. This process varies for different proteins, but fibronectin (FN) fibrillogenesis is well-characterized and serves as a model system for cell-mediated ECM assembly. Specifically, cells use integrin receptors on the cell membrane to bind FN dimers and actomyosin-generated contractile forces to unfold and expose binding sites for assembly into insoluble fibers. This receptor-mediated process enables cells to assemble and organize the ECM from the cellular to tissue scales. Here, we present a method termed surface-initiated assembly (SIA), which recapitulates cell-mediated matrix assembly using protein-surface interactions to unfold ECM proteins and assemble them into insoluble fibers. First, ECM proteins are adsorbed onto a hydrophobic polydimethylsiloxane (PDMS) surface where they partially denature (unfold) and expose cryptic binding domains. The unfolded proteins are then transferred in well-defined micro- and nanopatterns through microcontact printing onto a thermally responsive poly(N-isopropylacrylamide) (PIPAAm) surface. Thermally-triggered dissolution of the PIPAAm leads to final assembly and release of insoluble ECM protein nanofibers and nanostructures with well-defined geometries. Complex architectures are possible by engineering defined patterns on the PDMS stamps used for microcontact printing. In addition to FN, the SIA process can be used with laminin, fibrinogen and collagens type I and IV to create multi-component ECM nanostructures. Thus, SIA can be used to engineer ECM protein-based materials with precise control over the protein composition, fiber geometry and scaffold architecture in order to recapitulate the structure and composition of the ECM in vivo.

A method to obtain nanofibers and complex nanostructures from single or multiple extracellular matrix proteins is described. This method uses protein-surface interactions to create free-standing protein-based materials with tunable composition and architecture for use in a variety of tissue engineering and biotechnology applications.

细胞外基质蛋白纳米纤维和纳米结构工程采用表面引发大会

The extracellular matrix (ECM) in tissues is synthesized and assembled by cells to form a 3D fibrillar, protein network with tightly regulated fiber ...

Formation of Biomembrane Microarrays with a Squeegee-based Assembly Method

Lipid bilayer membranes form the plasma membranes of cells and define the boundaries of subcellular organelles. In nature, these membranes are heterogeneous mixtures of many types of lipids, contain membrane-bound proteins and are decorated with carbohydrates. In some experiments, it is desirable to decouple the biophysical or biochemical properties of the lipid bilayer from those of the natural membrane. Such cases call for the use of model systems such as giant vesicles, liposomes or supported lipid bilayers (SLBs). Arrays of SLBs are particularly attractive for sensing applications and mimicking cell-cell interactions. Here we describe a new method for forming SLB arrays. Submicron-diameter SiO2 beads are first coated with lipid bilayers to form spherical SLBs (SSLBs). The beads are then deposited into an array of micro-fabricated submicron-diameter microwells. The preparation technique uses a "squeegee" to clean the substrate surface, while leaving behind SSLBs that have settled into microwells. This method requires no chemical modification of the microwell substrate, nor any particular targeting ligands on the SSLB. Microwells are occupied by single beads because the well diameter is tuned to be just larger than the bead diameter. Typically, more 75% of the wells are occupied, while the rest remain empty. In buffer SSLB arrays display long-term stability of greater than one week. Multiple types of SSLBs can be placed in a single array by serial deposition, and the arrays can be used for sensing, which we demonstrate by characterizing the interaction of cholera toxin with ganglioside GM1. We also show that phospholipid vesicles without the bead supports and biomembranes from cellular sources can be arrayed with the same method and cell-specific membrane lipids can be identified.

Supported lipid bilayers and natural membrane particles are convenient systems that can approximate the properties of cell membranes and be incorporated in a variety of analytical strategies. Here we demonstrate a method for preparing microarrays composed of supported lipid bilayer-coated SiO2 beads, phospholipid vesicles or natural membrane particles.

生物膜的微阵列用刮墨刀型组合方式形成

Lipid bilayer membranes form the plasma membranes of cells and define the boundaries of subcellular organelles. In nature, these membranes are ...

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Metabolomics is a methodology used for the identification and quantification of many low-molecular-weight intermediates and products of metabolism within a cell, tissue, organ, biological fluid, or organism. Metabolomics traditionally focuses on water-soluble metabolites. The water-soluble metabolome is the final product of a complex cellular network that integrates various genomic, epigenomic, transcriptomic, proteomic, and environmental factors. Hence, the metabolomic analysis directly assesses the outcome of the action for all these factors in a plethora of biological processes within various organisms. One of these organisms is the budding yeast Saccharomyces cerevisiae, a unicellular eukaryote with the fully sequenced genome. Because S. cerevisiae is amenable to comprehensive molecular analyses, it is used as a model for dissecting mechanisms underlying many biological processes within the eukaryotic cell. A versatile analytical method for the robust, sensitive, and accurate quantitative assessment of the water-soluble metabolome would provide the essential methodology for dissecting these mechanisms. Here we present a protocol for the optimized conditions of metabolic activity quenching in and water-soluble metabolite extraction from S. cerevisiae cells. The protocol also describes the use of liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for the quantitative analysis of the extracted water-soluble metabolites. The LC-MS/MS method of non-targeted metabolomics described here is versatile and robust. It enables the identification and quantification of more than 370 water-soluble metabolites with diverse structural, physical, and chemical properties, including different structural isomers and stereoisomeric forms of these metabolites. These metabolites include various energy carrier molecules, nucleotides, amino acids, monosaccharides, intermediates of glycolysis, and tricarboxylic cycle intermediates. The LC-MS/MS method of non-targeted metabolomics is sensitive and allows the identification and quantitation of some water-soluble metabolites at concentrations as low as 0.05 pmol/&#181;L. The method has been successfully used for assessing water-soluble metabolomes of wild-type and mutant yeast cells cultured under different conditions.

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

We present a protocol for the identification and quantitation of major classes of water-soluble metabolites in the yeast Saccharomyces cerevisiae. The described method is versatile, robust, and sensitive. It allows the separation of structural isomers and stereoisomeric forms of water-soluble metabolites from each other.

使用液体色谱加上串联质谱法的 糖核 酸菌的定量元细胞学

Metabolomics is a methodology used for the identification and quantification of many low-molecular-weight intermediates and products of metabolism within ...

生物化学

Design and Implementation of an Automated Illuminating, Culturing, and Sampling System for Microbial Optogenetic Applications

Optogenetic systems utilize genetically-encoded proteins that change conformation in response to specific wavelengths of light to alter cellular processes. There is a need for culturing and measuring systems that incorporate programmed illumination and stimulation of optogenetic systems. We present a protocol for building and using a continuous culturing apparatus to illuminate microbial cells with programmed doses of light, and automatically acquire and analyze images of cells in the effluent. The operation of this apparatus as a chemostat allows the growth rate and the cellular environment to be tightly controlled. The effluent of the continuous cell culture is regularly sampled and the cells are imaged by multi-channel microscopy. The culturing, sampling, imaging, and image analysis are fully automated so that dynamic responses in the fluorescence intensity and cellular morphology of cells sampled from the culture effluent are measured over multiple days without user input. We demonstrate the utility of this culturing apparatus by dynamically inducing protein production in a strain of Saccharomyces cerevisiae engineered with an optogenetic system that activates transcription.

We designed a continuous culturing apparatus for use with optogenetic systems to illuminate cultures of microbes and regularly image cells in the effluent with an inverted microscope. The culturing, sampling, imaging, and image analysis are fully automated so that dynamic responses to illumination can be measured over several days.

设计和自动照明，培养和取样系统实现微生物光遗传学的应用

Optogenetic systems utilize genetically-encoded proteins that change conformation in response to specific wavelengths of light to alter cellular ...

Light-Controlled Fermentations for Microbial Chemical and Protein Production

Microbial cell factories offer a sustainable alternative for producing chemicals and recombinant proteins from renewable feedstocks. However, overburdening a microorganism with genetic modifications can reduce host fitness and productivity. This problem can be overcome by using dynamic control: inducible expression of enzymes and pathways, typically using chemical- or nutrient-based additives, to balance cellular growth and production. Optogenetics offers a non-invasive, highly tunable, and reversible method of dynamically regulating gene expression. Here, we describe how to set up light-controlled fermentations of engineered Escherichia coli and Saccharomyces cerevisiae for the production of chemicals or recombinant proteins. We discuss how to apply light at selected times and dosages to decouple microbial growth and production for improved fermentation control and productivity, as well as the key optimization considerations for best results. Additionally, we describe how to implement light controls for lab-scale bioreactor experiments. These protocols facilitate the adoption of optogenetic controls in engineered microorganisms for improved fermentation performance.

Optogenetic control of microbial metabolism offers flexible dynamic control over fermentation processes. The protocol here shows how to set up blue light-regulated fermentations for chemical and protein production at different volumetric scales.

用于微生物化学和蛋白质生产的光控发酵

Microbial cell factories offer a sustainable alternative for producing chemicals and recombinant proteins from renewable feedstocks. However, ...

Acquiring Fluorescence Time-lapse Movies of Budding Yeast and Analyzing Single-cell Dynamics using GRAFTS

Fluorescence time-lapse microscopy has become a powerful tool in the study of many biological processes at the single-cell level. In particular, movies depicting the temporal dependence of gene expression provide insight into the dynamics of its regulation; however, there are many technical challenges to obtaining and analyzing fluorescence movies of single cells. We describe here a simple protocol using a commercially available microfluidic culture device to generate such data, and a MATLAB-based, graphical user interface (GUI) -based software package to quantify the fluorescence images. The software segments and tracks cells, enables the user to visually curate errors in the data, and automatically assigns lineage and division times. The GUI further analyzes the time series to produce whole cell traces as well as their first and second time derivatives. While the software was designed for S. cerevisiae, its modularity and versatility should allow it to serve as a platform for studying other cell types with few modifications.

We present a simple protocol to obtain fluorescence microscopy movies of growing yeast cells, and a GUI-based software package to extract single-cell time series data. The analysis includes automated lineage and division time assignment integrated with visual inspection and manual curation of tracked data.

获取荧光时间推移电影的芽殖酵母和分析单细胞动力学移植物

Fluorescence time-lapse microscopy has become a powerful tool in the study of many biological processes at the single-cell level. In particular, movies ...

生物学

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

Cyanobacteria are a diverse group of prokaryotic photosynthetic organisms that can be genetically modified for the renewable production of useful industrial commodities. Recent advances in synthetic biology have led to development of several cloning toolkits such as CyanoGate, a standardized modular cloning system for building plasmid vectors for subsequent transformation or conjugal transfer into cyanobacteria. Here we outline a detailed method for assembling a self-replicating vector (e.g., carrying a fluorescent marker expression cassette) and conjugal transfer of the vector into the cyanobacterial strains Synechocystis sp. PCC 6803 or Synechococcus elongatus UTEX 2973. In addition, we outline how to characterize the performance of a genetic part (e.g., a promoter) using a plate reader or flow cytometry.

Here, we present a protocol describing how to i) assemble a self-replicating vector using the CyanoGate modular cloning toolkit, ii) introduce the vector into a cyanobacterial host by conjugation, and iii) characterize transgenic cyanobacteria strains using a plate reader or flow cytometry.

使用CyanoGate模块化克隆工具包进行共生对蓝藻的遗传改造

Cyanobacteria are a diverse group of prokaryotic photosynthetic organisms that can be genetically modified for the renewable production of useful ...

Isolation and Characterization of Intact Phycobilisome in Cyanobacteria

In cyanobacteria, phycobilisome is a vital antenna protein complex that harvests light and transfers energy to photosystem I and II for photochemistry. Studying the structure and composition of phycobilisome is of great interest to scientists because it reveals the evolution and divergence of photosynthesis in cyanobacteria. This protocol provides a detailed and optimized method to break cyanobacterial cells at low cost by a bead-beater efficiently. The intact phycobilisome can then be isolated from the cell extract by sucrose gradient ultracentrifugation. This method has demonstrated being suitable for both model and non-model cyanobacteria with different cell types. A step-by-step procedure is also provided to confirm the integrity and property of phycobiliproteins by 77K fluorescence spectroscopy and SDS-PAGE stained by zinc sulfate and Coomassie Blue. The isolated phycobilisome can also be subjected to further structural and compositional analyses. Overall, this protocol provides a helpful starting guide that allows researchers unfamiliar with cyanobacteria to quickly isolate and characterize intact phycobilisome.

The current protocol details the isolation of phycobilisomes from cyanobacteria by centrifugation through a discontinuous sucrose density gradient. The fractions of intact phycobilisomes are confirmed by 77K fluorescent emission spectrum and SDS-PAGE analysis. The resulting phycobilisome fractions are suitable for negative staining of TEM and mass spectrometry analysis.

蓝藻中完整藻胆体的分离与表征

In cyanobacteria, phycobilisome is a vital antenna protein complex that harvests light and transfers energy to photosystem I and II for photochemistry. ...

异养酵母与光养分泌糖的蓝细菌相结合的共培养物的组装和定量

摘要

探索更多视频

Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry

气泡柱光生物反应器中绿色微藻的培养及中性脂质的测定

细胞外基质蛋白纳米纤维和纳米结构工程采用表面引发大会

生物膜的微阵列用刮墨刀型组合方式形成

使用液体色谱加上串联质谱法的糖核酸菌的定量元细胞学

设计和自动照明，培养和取样系统实现微生物光遗传学的应用

用于微生物化学和蛋白质生产的光控发酵

获取荧光时间推移电影的芽殖酵母和分析单细胞动力学移植物

使用CyanoGate模块化克隆工具包进行共生对蓝藻的遗传改造

蓝藻中完整藻胆体的分离与表征

异养酵母与光养分泌糖的蓝细菌相结合的共培养物的组装和定量

摘要

探索更多视频

Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry

气泡柱光生物反应器中绿色微藻的培养及中性脂质的测定

细胞外基质蛋白纳米纤维和纳米结构工程采用表面引发大会

生物膜的微阵列用刮墨刀型组合方式形成

使用液体色谱加上串联质谱法的 糖核 酸菌的定量元细胞学

设计和自动照明，培养和取样系统实现微生物光遗传学的应用

用于微生物化学和蛋白质生产的光控发酵

获取荧光时间推移电影的芽殖酵母和分析单细胞动力学移植物

使用CyanoGate模块化克隆工具包进行共生对蓝藻的遗传改造

蓝藻中完整藻胆体的分离与表征

使用液体色谱加上串联质谱法的糖核酸菌的定量元细胞学