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Past Issues in 2019

Volume: 9 Issue: 7

Microbiology

Delivering "Chromatic Bacteria" Fluorescent Protein Tags to Proteobacteria Using Conjugation Authors: Rudolf O Schlechter and Mitja NP Remus-Emsermann, date: 04/05/2019, view: 4681, Q&A: 0

Recently, we published a large and versatile set of plasmids, the chromatic bacteria toolbox, to deliver eight different fluorescent protein genes and four combinations of antibiotic resistance genes to Gram-negative bacteria. Fluorescent tags are important tools for single-cell microbiology, synthetic community studies, biofilm, and host-microbe ...

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Quantification of Hepatitis B Virus Covalently Closed Circular DNA in Infected Cell Culture Models by Quantitative PCR Authors: Bingqian Qu and Stephan Urban, date: 04/05/2019, view: 4251, Q&A: 0

Persistence of the human hepatitis B virus (HBV) requires the maintenance of covalently closed circular (ccc)DNA, the episomal genome reservoir in nuclei of infected hepatocytes. cccDNA elimination is a major aim in future curative therapies currently under development. In cell culture based in vitro studies, both hybridization- and ...

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Multiple Modification of Chromosomal Loci Using URA5.3 Selection Marker in the Unicellular Red Alga Cyanidioschyzon merolae Authors: Tokiaki Takemura, Sousuke Imamura, Yuki Kobayashi and Kan Tanaka, date: 04/05/2019, view: 3398, Q&A: 0

The unicellular red alga Cyanidioschyzon merolae has been used as a eukaryotic photosynthetic model for various basic and applied studies. Although the nuclear genome of C. merolae can be modified by homologous recombination with exogenously introduced DNA, it has been difficult to modify multiple chromosome loci within the same ...

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Molecular Biology

Identification of RNase-sensitive LINE-1 Ribonucleoprotein Interactions by Differential Affinity Immobilization Authors: Hua Jiang, Martin S. Taylor, Kelly R. Molloy, Ilya Altukhov and John LaCava, date: 04/05/2019, view: 4149, Q&A: 0

Long Interspersed Nuclear Element-1 (LINE-1, L1) constitutes a family of autonomous, self-replicating genetic elements known as retrotransposons. Although most are inactive, copious L1 sequences populate the human genome. L1s proliferate in a ‘copy-and-paste’ fashion through an RNA intermediate; a full-length L1 transcript is ~6,000 nucleotides ...

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Neuroscience

Consummatory Successive Negative Contrast in Rats Authors: Ana María Jiménez-García, Leandro Ruiz-Leyva, Ana Vázquez-Ágredos, Carmen Torres, Mauricio R. Papini, Cruz Miguel Cendán and Ignacio Morón, date: 04/05/2019, view: 2819, Q&A: 0

Using animal models in addiction and pain research is pivotal to unravel new pathways and mechanisms for the treatment of these disorders. Reward devaluation through a consummatory successive negative contrast (cSNC) task has shown the ability to reduce physical pain sensitivity (hypoalgesia) and increase oral ethanol consumption in rats. The ...

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Plant Science

Quantification of the Humidity Effect on HR by Ion Leakage Assay Authors: Musoki Mwimba and Xinnian Dong, date: 04/05/2019, view: 3175, Q&A: 0

We describe a protocol to measure the contribution of humidity on cell death during the effector-triggered immunity (ETI), the plant immune response triggered by the recognition of pathogen effectors by plant resistance genes. This protocol quantifies tissue cell death by measuring ion leakage due to loss of membrane integrity during the ...

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Visualization of Plant Cell Wall Epitopes Using Immunogold Labeling for Electron Microscopy Author: Mateusz Majda, date: 04/05/2019, view: 4102, Q&A: 0

Plant cell walls consist of different polysaccharides and structural proteins, which form a rigid layer located outside of the plasma membrane. The wall is also a very dynamic cell composite, which is characterized by complex polysaccharide interactions and various modifications during cell development. The visualization of cell wall components in ...

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Quantitative Plasmodesmata Permeability Assay for Pavement Cells of Arabidopsis Leaves Authors: Min Diao, Qiannan Wang and Shanjin Huang, date: 04/05/2019, view: 4168, Q&A: 0

Plasmodesmata (PD) are intercellular channels between walled plant cells that enable the transportation of materials between adjacent cells, which are important for plant growth and development. The permeability of PD must be tightly regulated. Assays to determine the permeability of PD are crucial for related studies on the regulation of PD ...

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Extraction and Purification of Laccases from Rice Stems Authors: Chenna Swetha and P. V. Shivaprasad, date: 04/05/2019, view: 3583, Q&A: 0

Laccases are found in cell walls of plants in very low amounts. This protocol provides an efficient method to purify laccases from rice stems. The method involves three steps: 1) Isolation of total protein from rice stems using buffers with high salt concentration to extract protein from cell walls; 2) Purification of laccases using concanavalin-A ...

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