Extraction of proteins from wood samples colonized by bioprotectants and/or stain fungi

  • 74 Pages
  • 3.86 MB
  • English
Plant proteins -- Diagnostic use -- Technique., Plant proteins -- Separation., Blue stain -- Biological control -- Testing., Ponderosa pine -- Diseases and pests -- Biological control -- Tes
Statementby Li Cui.
The Physical Object
Pagination74 leaves, bound ;
ID Numbers
Open LibraryOL15445311M

Extraction of proteins from wood samples colonized by a technique was identified to extract proteins from ponderosa pine sapwood samples colonized by bioprotectants and/or sapstain fungi. These extracts could be used to study the effects of the bioprotectants on various target stain fungi.

The effects of extraction media and sample Author: Li Cui. EXTRACTION OF PROTEINS FROM WOOD SAMPLES COLONIZED BY BIOPROTECTANTS AND/OR STAIN FUNGI 1 INTRODUCTION Sapstain of wood (also called blue stain) is a common problem in both softwoods and hardwoods.

Most sapstains are caused by fungi and bacteria. Stained wood is not accepted by customers and losses due to reduced marketability of the discolored.

Description Extraction of proteins from wood samples colonized by bioprotectants and/or stain fungi FB2

Protein recovery and\ud enzyme activity were affected by the extraction time, the\ud medium conditions of the bioprotectants and the sample\ud incubation conditions.

The greatest protein recovery\ud occurred, under the conditions of this study, when wood\ud samples were incubated for thirty days and extracted for\ud twelve hours using 50 mM. Summary We investigated the ability of selected buffers to extract proteins and other materials from the surfaces and subsurfaces of ponderosa pine wafers colonized by selected wood stain fungi and potential bioprotectant bacteria.

The effects of extraction conditions on enzyme activity were also analyzed. The addition of Tween 80 to the extraction media markedly enhanced total protein. In this chapter we present a protocol for total protein extraction optimized for wood-forming tissue (differentiating secondary xylem).

The protocol is then used for a series of other organs (root, leaf, pollen, bud, flower, cambium, and phloem) in broadleaf (oak and poplar) and conifer (pine) by: 6. Extraction of proteins from environmental samples Ten milliliters of wastewater, or 1 g each of soil or sediment were used as starting material for direct extraction of proteins.

The boiling method. For wastewater, microorganisms and other particulate matter were collected from the aqueous phase by centrifugation at 12 x g for 10 min. Extraction of proteins from wood samples colonized by bioprotectants and/or stain fungi samples colonized by bioprotectants and/or sapstain fungi.

These extracts could be used to study the. Hence, a common protein extraction protocol is desirable. In the present study, soluble proteins were extracted from six diverse samples using TRIzol without any additional clean-up step and subjected to 2-DE and 2D-DIGE analysis for global protein expression profiling.

Image analysis using IMP7 and DeCyder showed good coverage, reproducibility. This handbook is divided into five chapters, starting with an overview of protein sample preparation, followed by three chapters based on a conceptual, high-level workflow for protein sample preparation and analysis (Fig I.1), and wrapping up with a chapter that focuses on.

Proteins (also known as polypeptides) are organic compounds made of amino acids arranged in a linear chain. The amino acids in a polymer are joined together by the peptide bonds between the carboxyl and the amino groups of adjacent amino acid residues.

Like other biological macromolecules such as polysaccharides and nucleic acids. Li Cui MSc Fall ‘95 (Extraction of proteins from wood samples colonised by bioprotectants and/or stain fungi) Lori Elkins MSc Summer ‘05 (co w/ R.

Leichti) (Establishing a through-boring pattern and method of test for utility poles). Peroxide post-treatment of wood pressure impregnated with MBCC was shown to reduce color change by more than 50% compared to controls.

Erosion due to photo-degradation and colonization by black-stain fungi were lower in samples treated with MBCC than in untreated controls and were relatively unaffected by peroxide post-treatment.

Sigma-Aldrich has developed protein extraction kits that are specialized for the extraction and isolation of high quality protein from plant tissue samples. Kits contain a plant specific protease inhibitor cocktail and plant specific extractions reagents.

The protocols and the extraction reagents are optimized to significantly remove interfering substances such as tannins, phenolics, and flavones. Sample Preparation. Maize-leaf tissue was ground to a fine powder with mortar and pestle in liquid nitrogen and lyophilized. Twelve tubes, each one with an equivalent of mg of fresh weight (FW), corresponding to 30 mg of dry powder, were prepared and used to extract protein in triplicate, using five different methods.

Detector register the light that pass trough the sample (↑ absorption of light - ↑ concentration of nucleic acids) DNA/RNA is present Results can be altered by contaminants (phenol, proteins) / = stable / = o > (contamination with proteins) / = o.

DNA was isolated from pure cultures of these fungi and also from spruce wood blocks colonized by individual isolates of wood decay basidiomycetes or wood-inhabiting ascomycetes.

The primer pair ITS1-F (specific for higher fungi) and ITS4 (universal primer) amplified the internal transcribed spacer region from both ascomycetes and basidiomycetes. A rapid procedure for protein extraction from filamentous fungi and plants using YeastBuster™ Protein Extraction Reagent 59 Double your chance to discover less-abundant, low-solubility proteins on 2D electrophoresis gels 61 Convenient and versatile subcellular extraction procedure that facilitates classical protein expression.

Details Extraction of proteins from wood samples colonized by bioprotectants and/or stain fungi PDF

The recommended minimum sample for extraction can be as little as g tissue or million cells, but in certain cases using less reagent can be effective with an even smaller sample. The Total Protein Extraction Kit comes with a protease inhibitor cocktail to prevent protein degradation.

Figure 1. Protein Extraction Beads are required for efficient tissue disruption using the Bioruptor® Plus Complete disruption is observed in the sample containing Diagenode’s Protein Extraction Beads (left) after 5 cycles while non-disrupted tissue is still present in the sample without the Protein Extraction.

Enough Coomassie Stain (EzBlue R stain reagent Sigma Aldrich) was added to cover the gel by 1/2 inch (∼ cm) and incubated for 1 h on gel rocker The solution was decanted and gel was washed with distilled-water for 2–3 intensify protein bands gel was incubated in destaining solution (methanol −30%, acetic acid −10% in water.

Supplemental methods for protein extraction and western blot. Method used for protein extraction, western-blot analysis, production and purification of 6His-PtXLIM1a recombinant protein.

Download Extraction of proteins from wood samples colonized by bioprotectants and/or stain fungi FB2

View. MicroRotofor lysis kits provide cell lysis and protein extraction protocols that are tailored to the specific needs of different sample sources. ReadyPrep™ Mini Grinders ReadyPrep mini grinders are used in sample extraction protocols to grind small biological samples for high recovery of proteins.

Protein fusion tags are used to aid expression of suitable levels of soluble protein as well as purification. A unique protein tag, the HaloTag® protein, is engineered to enhance expression and solubility of recombinant proteins in E. coli. HaloTag® protein tag is a 34kDa, monomeric protein tag modified from Rhodococcus rhodochrous dehalogenase.

The advent of recombinant DNA technology and the overexpression of heterologous proteins in bacteria have posed some unique problems not previously encountered in extraction of bacterial proteins.

Some of this technology has enabled secretion of recombinant proteins by bacteria into the media, thereby eliminating the need to lyze the cells. EXTRACTION AND CHARACTERIZATION OF PROTEINS Abstract Different techniques and principles for protein extraction and characterization were demonstrated in this experiment.

Various proteins were extracted from different sources: g yeast invertase, g egg white albumin, and g of milk casein. Activity assay for invertase was performed using Benedict’s test and the. Protein extraction and sample cleanup are the most important steps to ensure optimal resolution and reduce variability of your 2-D gels.

2-D PAGE success depends on sample purity. Interfering substances that can negatively impact SDS-PAGE and 2-DGE include salts, detergents, denaturants, or organic solvents, so it is crucial to eliminate these. Plant tissues contain large amounts of secondary compounds that significantly interfere with protein extraction and 2DE analysis.

Thus, sample preparation is a crucial step prior to 2DE in plant proteomics. This tutorial highlights the guidelines that need to be followed to perform an adequate total protein extraction before 2DE in plant. Spin down samples for 15 min at 20, x g.

Three phases are visible after centrifugation as shown in the protein extraction flow chart below. Carefully aspirate the middle phase of sample extract using a micropipette with a 20‐ l range (e.g. Gilson P).

Often plant tissues are recalcitrant and, due to that, methods relying on protein precipitation, such as TCA/acetone precipitation and phenol extraction, are usually the methods of choice for protein extraction in plant proteomic studies.

However, the addition of precipitation steps to protein extraction methods may negatively impact protein recovery, due to problems associated with protein. Protein-based biopolymers, such as zein (a hydrophobic protein extracted from maize), and gelatin (a mixture of animal peptides and proteins obtained by the partial hydrolysis of collagen) can be efficiently used for wood surface protection, mainly when an ionic liquid (1-ethylmethylimidazolium chloride) is used as solvent-carrier (Croitoru.

Proteomics14, – DOI /pmic TUTORIAL Protein extraction from plant tissues for 2DE and its application in proteomic analysis Xiaolin Wu, Fangping Gong and Wei Wang State Key Laboratory of Wheat & Maize Crop Science in .Proteins secreted by plant cells into the extracellular space, consisting of the cell wall, apoplastic fluid, and rhizosphere, play crucial roles during development, nutrient acquisition, and stress acclimation.

However, isolating the full range of secreted proteins has proven difficult, and new strategies are constantly evolving to increase the number of proteins that can be detected and.

Principles and Reactions of Protein Extraction, Purification, and Characterization provides the mechanisms and experimental procedures for classic to cutting-edge techniques used in protein extraction, purification, and characterization.

The author presents the principles and reactions behind each procedure and uses tables to compare the different.