Protoplast isolation from tomato - posted in Botany and Plant Biology: I am trying to isolate protoplast from tomato, exactly the way Jen Sheens lab does for Arabidopsis, but the yield is very very poor. Moreover I know that eventually all I see after isolation is collection of chloroplast. I think that the osmotic pressure is very to disturbed. Does anybody have a set of protocol to isolate protoplast from tomato. Kindly suggest.
Transient assays using protoplasts are ideal for processing large quantities of genetic data coming out of hi-throughput assays. Previously, protoplasts have routinely been prepared from dicot tissue or cell suspension cultures and yet a good system for rice protoplast isolation and manipulation is lacking.. We have established a rice seedling protoplast system designed for the rapid characterization of large numbers of genes. We report optimized methods for protoplast isolation from 7-14 day old etiolated rice seedlings. We show that the reporter genes luciferase GL2 and GUS are maximally expressed approximately 20 h after polyethylene glycol (PEG)-mediated transformation into protoplasts. In addition we found that transformation efficiency varied significantly with plasmid size. Five micrograms of a 4.5 kb plasmid resulted in 60-70% transformation efficiency. In contrast, using 50 microg of a 12 kb plasmid we obtained a maximum of 25-30% efficiency. We also show that short interfering RNAs ...
Plant transformation and exogenous protein expression is essential for molecular biology and biotechnology. Current approaches of stable plant transformation might be problematic and very time-consuming. Because of this, transient expression in protoplasts has become valuable alternative, being less cost and time-effective at the same time. Excellent for eukaryotic proteins, representing a natural cell habitat, protoplast isolation is widely used in protein interaction visualization techniques, like BiFC (Bimolecular fluorescence complementation) and FRET (Förster resonance energy transfer). In this protocol we present a another use of Arabidopsis protoplast in protein degradation assay, proving its high versatility as a tool in proteomics.
Explant-dependent receptivity to isolation and a cell-wall resynthesis in protoplast culture of recalcitrant yellow lupin. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
SUMMARY: Conditions for highly efficient genetic recombination in Streptomyces by protoplast fusion are described. Protoplasts of S. fradiae and S. griseofuscus were formed by a modification of the glycine-lysozyme-lytic enzyme method (Okanishi, Suzuki & Umezawa, 1974). Regeneration of cells from protoplasts was monitored throughout the growth cycle and was most efficient when cells of either S. fradiae or S. griseofuscus were taken from the transition phase between the exponential and stationary growth phases. Fusion of protoplasts carrying different auxotrophic or chromosomal drug-resistance markers was achieved by treatment with polyethylene glycol, and high frequencies of stable genetic recombinants were obtained.
Guard cell protoplasts (GCPs) were isolated from the adaxial epidermis of Vicia leaves. The properties of isolated adaxial GCPs (ad GCPs) were compared with tho
Uptake and metabolism of exogenous naphthalene-1-acetic acid (NAA) and indole-3-acetic acid (IAA) have been studied in tobacco (Nicotiana tabacum L. cv. Xanthi) mesophyll protoplasts. Both auxins entered protoplasts by diffusion under the action of the transmembrane pH gradient without any detectable participation of an influx carrier. Molecules were accumulated by an anion-trapping mechanism and most of them were metabolized within hours, essentially as glucose-ester and amino-acid conjugates. Protoplasts were equipped with a functional auxin-efflux carrier as evidenced by the inhibitory effect of naphthylphtalamic acid on IAA efflux. Basically, similar mechanisms of NAA and IAA uptake occurred in protoplasts. However, the two auxins differed in their levels of accumulation, due to different membrane-transport characteristics, and the nature of the metabolites produced. This shows the need to estimate the accumulation and the metabolism of auxins when analyzing their effects in a given cell system. The
Author】 ZHANG Xiao-li1, HE Rui1*, TONG Jia-yun1,2, ZHAN Ruo-ting1, LIN Chao1 1. Key Laboratory of Chinese Medicinal Resources from Lingnan of Ministry of Education, Research Centre of Chinese Herbal Resource Science and Engineering, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; 2. College of Chinese Traditional Medicine,Guangzhou University of Chinese Medicine, GuangZhou, Guangdong 510006, China. 【摘要】 [Objective] To set up protoplast preparation and purification method from Nervilia fordii (Hance) Schltr. leaves. [Method] Taking the yield and survival rate of protoplast as indices, the effects of different factors, including pretreatment, enzymolysis method, enzymolysis temperature, centrifugation speed, illumination, and sieve mesh number were studied for the separation and purification of protoplast from N. fordii leaves. [Result] The optimum process for protoplast isolation was obtained as follows: N. fordii leaves were first pretreated for 1 h in CPW ...
SUMMARY: Protoplasts of Saccharomyces cerevisiae cultivaued in sporulation medium with an osmotic stabilizer at pH 7.0 were studied by means of light and electron microscopy. After 15 h, mature ascospores with complese cell walls were formed in the protoplasts. During sporulation, protoplasts regenerated only the fibrillar wall component and were osmotically sensitive. By using snail enzymes to block fibril regeneration on the protoplast surface, it was proved that sporulation was quite independent of such regeneration. Most of the ascospores produced by protoplasts were viable, osmotically resistant and save rise to new yeast cells. Meiotic division and sporulation in the yeasts are processes quite independent of the presence of the ascus wall.
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Among the eight strains of Listeria monocytogenes tested for lysozyme sensitivity, two were resistant to lysozyme but became sensitive after lipase pretreatment. Among the other six, one was very sensitive to lipase and another one was extremely susceptible to lysozyme. Stable protoplasts were formed from the lysozyme-resistant strain (42) by lipase and lysozyme treatment, which completely digested the cell wall. The cell wall (uranyl acetate-lead stained) was of a thick triple-layered profile, with the intermediate layer of low density. Lipase treatment for a short time (60 min) did not cause any alteration in structure, but prolonged treatment (180 min) caused extensive digestion of the plasma membrane and the cell wall, liberating cytoplasmic material. When the cells were treated with either lipase or lysozyme, a small number of protoplasts were extruded through the partly digested or weakened transverse cell wall, leaving an almost intact cell wall ghost. There were small vesicular structures in the
106. Abstract Protoplasts of Boergesenia forbesii (Hanvey) were treated with inhibitors of protein synthesis in order to investigate their effects on cellulose synthesis. Cellulose synthesis was reversibly inhibited by 10 M cycloheximide as assayed by fluorescence microscopy of Tinopal binding to cellulose. Freeze fracture and image analysis of cycloheximide-treated cells indicated a reduction in the number of intramembrane particles; however, the terminal synthesizing complexes remained at all times. Treatment with 10 M actinomycin D, when applied during the first hour of protoplast formation, irreversibly inhibits cellulose synthesis and terminal complex formation. De novo protein synthesis is required for cell wall regeneration by protoplasts. The data suggest that the structural subunits visualized in the terminal complex do not undergo signifi-cant turnover, but that there may exist an essential proteinaceous component of cellulose synthesis which must be continually renewed ...
Light micrograph of protoplasts from a tobacco leaf, Nicotiana plumbaginifolia, showing two different cell types, epidermal (colourless) & mesophyll (green). A protoplast is a single intact cell, which has been chemically treated to remove the tough outer cellulose wall, leaving only the delicate plasma membrane binding the cell contents into a whole. The cell wall can later be regrown. The epidermal cells, a layer at the surface, protect the leaf from external damage. The mesophyll, a layer beneath, contain numerous chloroplasts, sites of photosynthesis. Protoplast cells, being easier to manipulate, are used in plant genetic experimentation. Mag: X125 (35mm). - Stock Image B060/0002
1) Two extracellular staphylolytic enzymes have been Isolated and purified from submerged cultures of Streptomyces griseus. 2) The enzymes were electrophoretically homogeneous at several pH values. A single peak was obtained for each protein on ultracentrifugation. Based on gel-filtration molecular weights of 11,800 and 13,000 were determined for enzyme 1 and enzyme 2 respectively. 3) The ionic strength and pH optima for lytic activity of the two enzymes against staphylococcal cells and their isolated cell walls have been detenained. The effects of several cations and certain known enzyme inhibitors have been tested on lytic action of the two enzymes against cell walls. 4) Both enzymes were N-acetylhexosaminidases. Examination of the acid hydrolysis products of fully digested cell walls treated with naBH4 demonstrated that both enzymes were muramidases. 5) Both enzymes caused lysis to varying extents, of the cell walls of a range of gram positive organisms. However, E. coli, the only gram ...
Sigma-Aldrich offers abstracts and full-text articles by [Marco Ieronimo, Sergii Afonin, Katja Koch, Marina Berditsch, Parvesh Wadhwani, Anne S Ulrich].
A process for the production of aminoglycoside antibiotics using fused protoplasts derived from streptomyces. The cells are precultured in a medium containing sucrose, calcium and magnesium salts. Protoplasts are formed and then fused. The fused protoplasts are regenerate and antibiotic producing ability is screened for. The regenerate cells produced an antibiotic complex of modified composition or demonstrated increased or reduced antibiotic productivity compared to the fusion partners.
soft agar (0.8%) plates containing In some cases, protoplasts were resus- pended in LB medium containing 0.5 M sucrose and spread onto soft agar plates containing LB medium and 0.5 M sucrose with autoclaved plastic spreaders. The plates were subsequently incubated at either 25 or 37 1C. In other cases, the protoplasts were diluted into molten soft agar at 45 1C containing either LB medium and 0.5 M sucrose or M9 medium, 0.5 M sucrose, and either Leu (100 mg/ml) or Arg (100 mg/ml), depending on the nutritional requirements of the auxotrophic strains used, and then poured onto soft agar plates of the same composition. ...
Auxin modifies the electrical transmembrane potential of isolated protoplasts according to an inverted bell-shaped dose-response curve (Ephritikhine et al, 1987; Shen et al, 1988). The sensitivity of...
Abundant microtubules (MTs) were present both in protoplasts isolated from tobacco BY-2 cells and on membrane ghosts prepared from such protoplasts. However, only a few MTs or none at all were observed on membrane ghosts prepared from protoplasts pretreated with protease, trypsin or chymotrypsin, although abundant MTs were present in protease-pretreated protoplasts. These observations suggest that the digestion of the extracellular portion of transmembrane protein(s) results in the dissociation of cortical MTs from the plasma membrane. Exogenously applied extensin or poly-L-lysine increased the cold-stability of cortical MTs in the control protoplasts, but not in protease-pretreated protoplasts. It appears that the transmembrane protein(s) is also involved in the stabilization of cortical MTs by extensin or poly-L-lysine. Cortical MTs in BY-2 cells were arranged parallel to one another and were resistant to cold. Treatment with protease rendered the MTs sensitive to cold and often disturbed the ...
Plant, bacterial or fungal protoplasts & spheroplasts: what are the key differences, how are they prepared and what are they ultimately used for?
Automated isolation of protoplasts from plant cells with the ALS CellCelector for downstream processes like single cell RNA analysis
The data presented in this paper show that second messengers such as cGMP and cAMP can provoke [Ca2+]cyt elevations in tobacco protoplasts. In the case of cAMP and cGMP, the targets of action are likely to be intracellular, because even though membrane-permeable analogs at concentrations as low as 0.1 μm provoked a large [Ca2+]cyt response (Fig.1, A and B), membrane-impermeable cAMP and cGMP did not produce any significant effect (Fig. 6). An intracellular site of action is also supported by the fact that a significant [Ca2+]cyt response was induced by db-cGMP and db-cAMP in the absence of external Ca2+. Protoplasts in EGTA (zero external Ca2+) still showed [Ca2+]cyt responses to these second messengers, implicating the release of Ca2+ from internal stores as a mechanism for this response. Although variability was seen between protoplast preparations, cyclic-nucleotide-mediated [Ca2+]cyt increases were generally higher in the presence of external Ca2+. This suggests that cyclic mononucleotides ...
Consider a water balloon inside a cardboard box. This is a good model for a plant cell. Thus, there are two major components to a plant cell: (1) the box which is analogous to the cell wall, and (2) the water balloon which is analogous to the cytoplasm or protoplast. The protoplast is everything inside the cell wall. It is the living part of the cell and includes the cytoplasm, nucleus, vacuole, and other goodies. Like a cardboard box, the cell wall is relatively rigid, it is non-living and highly structured. Its more obvious functions are to support and protect the cell. It is produced by the protoplast.. Pores or air spaces (called intercellular spaces) exist between adjacent cells because of the difficulty of packing of cells with rigid walls. Try and squeeze a bunch of irregularly boxes into a room without leaving any space between them! The intercellular spaces are important for gas exchange and water transport, some movements (i.e., sensitive plants - water moves into/out of theses ...
Using freshly isolated maize mesophyll protoplasts and a transient expression method, I showed that the transcriptional activity of seven maize photosynthetic gene promoters is specifically and coordinately repressed by the photosynthetic end products sucrose and glucose and by the exogenous carbon source acetate. Analysis of deleted, mutated, and hybrid promoters showed that sugars and acetate inhibit the activity of distinct positive upstream regulatory elements without a common consensus. The metabolic repression of photosynthetic genes overrides other forms of regulation, e.g., light, tissue type, and developmental stage. Repression by sugars and repression by acetate are mediated by different mechanisms. The identification of conditions that avoid sugar repression overcomes a major obstacle to the study of photosynthetic gene regulation in higher plants.. ...
Bargmann, Bastiaan O.R., and Kenneth D. Birnbaum. Positive Fluorescent Selection Permits Precise, Rapid, and In-Depth Overexpression Analysis in Plant Protoplasts. Plant Physiology 149.3 (2009): 1231-1239. Web. 29 Nov. 2020. ...
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DNA-free gene editing can be achieved by using purified Cas9 enzymes with gRNA and transfecting them directly into your cells or protoplasts of interest.
The freeze-fracture morphology of the plasma membrane of cells and isolated protoplasts of plant callus suspensions has been investigated. Plasmolysis of suspension cells leads to the formation of 2 types of hexagonal arrays of intramembrane particles situated on the inner fracture face (PF). These arrays are interpreted as proteins that have crystallized in the plane of the membrane as the area of surrounding lipid bilayer is reduced during protoplast retraction from the cell wall. Time-course studies have revealed no positive relationship between the distribution of hexagonal arrays and the occurrence of microfibrils regenerated around isolated protoplasts during periods of culture. No evidence for the specialized transport functions attributed to hexagonal arrays of plant cells by previous workers has been found. ...
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The location of coat protein of alfalfa mosaic virus (AIMV) strain 425 was determined in protoplasts isolated from infected tobacco leaves and in in vitro inoculated tobacco protoplasts, using immunocytochemistry on ultrathin frozen sections labeled with colloidal gold. In infected tobacco leaves 5 days postinoculation (p.i.) coat protein is present ... read more in the cytoplasm and nucleus, especially around the nucleolus. In in vitro inoculated tobacco protoplasts coat protein was not present in the nucleus 6 hr p.i. These results indicate that the presence of coat protein in the nucleus is not necessary for viral replication. However, coat protein could be detected in the nucleus 48 hr p.i. Probably coat protein or virus particles accumulate in the nucleus late in infection. Minus-strand RNA, as part of the replication complex, could be detected in a 650 g pellet fraction of infected tobacco leaves but not in the nucleus, suggesting that replication of AIMV occurs outside the nucleus. show ...
A variety of red algal species have been identified as potential food sources for juvenile Greenlip Abalone, Haliotis laevigata (,5 mm shell length). To provide the red algal species in a diet suitable for juvenile abalone three propagation methods; spore production, protoplast isolation, and fragment culture were investigated. The potential algae requirements and consumable costs for each propagation method were determined, using experimental data and values from the literature, to assess the viability of utilizing each of these propagation methods in a commercial abalone nursery. The use of red algal spores required 592-52,000 kg of algae, depending on the level of spore release and the percentage of fertile algal thalli collected. Protoplast isolation reduced the amount of algal biomass to 8.55-910 kg but was affected by the efficiency of the isolation procedure. Even at an efficient production of 1 × 108 protoplasts·g-1 wet weight alga the cost of consumables (enzymes) was $US 13,576. A ...
Morphogenesis-Protoplast Culture: Questions 1-3 of 3. Get to the point IFS (Forests Services) Botany (Mains) questions for your exams.
Since the development of genome editing tools like CRISPR/Cas9, it is possible to modify the sequences of genes in a very specific manner. The molecular delivery into plant protoplasts to improve the quality of agricultural crops represents a major bottleneck in the routine application of CRISPR/Cas9 in modern plant breeding. To approach this need, we suppose using gold nanoparticle mediated (GNOME) laser transfection for delivery of CRISPR/Cas9 ribonucleoproteins (RNP) into potato protoplasts with high-throughput. As a proof-of-concept, we aim to reduce the toxic steroidal glykoalkaloid α-solanine in potatoes. GNOME laser transfection utilizes a picosecond Nd:YAG laser operating at 532 nm to excite surface plasmon resonance of membrane-attached gold nanoparticles. The strong absorption of laser light results in a temperature increase, leading to vaporization of the surrounding medium and to the formation of cavitation bubbles, which causes a transient permeabilization of the cell membrane. The ...
Dear colleagues! I am wondering if there is any rules or restrictions for vectors, used in electroporation of protoplasts. Can the same vector used for Agro-mediated transformation be used for protoplasts electroporation? Thanks in advance, Nikolai __________________________________ Ph.D.c. Nikolai Scherbak Örebro University 701 82 Örebro, SWEDEN Tel. +46-19-301034 Fax. +46-19-303566 email: nikolai.scherbak at nat.oru.se ...
Rapid transient elevation of cytoplasmic calcium (Ca2+) levels in plant cells is an early signaling event triggered by many environmental cues including abiotic and biotic stresses. Cellular Ca2+ levels and their alterations can be monitored by genetically encoded reporter systems such as the bioluminescent protein, aequorin. Employment of proteinaceous Ca2+ sensors is usually performed in transgenic lines that constitutively express the reporter construct. Such settings limit the usage of these Ca2+ biosensors to particular reporter variants and plant genetic backgrounds, which can be a severe constraint in genetic pathway analysis. Here we systematically explored the potential of Arabidopsis thaliana leaf mesophyll protoplasts, either derived from a transgenic apoaequorin-expressing line or transfected with apoaequorin reporter constructs, as a complementary biological resource to monitor cytoplasmic changes of Ca2+ levels in response to various biotic stress elicitors. We tested a range of ...
Aluminium (Al), a non-essential metal widespread in the environment that is known to be toxic to humans as well as to plants, can cause damage not only to the roots but also to the aerial parts of plants. Its toxicity has been recognized as one of the major factors that limit crop production on acid soil. Alternative oxidase, the respiratory terminal oxidase in plants, which contributes to maintain the electron flux and reduce mitochondrial ROS levels, is often dramatically induced to make plants to adapt better to stress conditions like Al stress. In this protocol, the expression of alternative oxidase induced by Al treatment was detected in Arabidopsis protoplasts using an adaptation of previous methods (Yamamoto et al., 2002; Li et al., 2011; Liu et al., 2014), which contribute to research on the mechanism of alternative oxidase in Al treatment.
A key to successful genome editing is efficient delivery of genetic materials, and it has been the main bottleneck in transformation of microalgae. In general, the cell wall is considered as the most significant barrier for the introduction of macromolecules into plant cells [16]. To avoid this problem, the cell wall is removed, and the resulting protoplasts are subjected to PEG-mediated transfection, which appears to be very effective without the need for expensive supplies and equipment [18, 30, 96]. With the proper removal of the cell wall, this technique can result in a transfection efficiency of up to 70% in cassava mesophyll protoplasts [97], which may offer an opportunity to improve microalgal transformation. In fact, as summarized in Table 1, there have been a few reports on PEG-mediated transformation of microalgae including Chlorella [26, 27, 29] and Cyanidioschyzon [28]. These attempts did not result in a greatly improved transformation efficiency; however, this technique can be ...
Hibiscus latent Singapore virus (HLSV) is a member of Tobamovirus and its full-length cDNA clones were constructed. The in vitro transcripts from two HLSV full-length cDNA clones, which contain a hepta-adenosine stretch (pHLSV-7A) and an octo-adenosine stretch (pHLSV-8A), are both infectious. The replication level of HLSV-7A in Nicotiana benthamiana protoplasts was 5-fold lower, as compared to that of HLSV-8A. The replicase proteins of HLSV-7A were produced through programmed -1 ribosomal frameshift (-1 PRF) and the 7A stretch was a slippery sequence for -1 PRF. Mutations to the downstream pseudoknot of 7A stretch showed that the pseudoknot was not required for the frameshift in vitro. The stretch was found to be extended to 8A after subsequent replication cycles in vivo. It is envisaged that HLSV employs the monotonous runs of A and -1 PRF to convert its 7A to 8A to reach higher replication for its survival in plants. © 2013 Elsevier Inc ...
The technique is highly dynamic and could be varied to suit the particular type of cell cultureâ ¦ The techniques include culture of cells, anthers, ovules and embryos on experimental to industrial scales, protoplast isolation and fusion, cell selection and meristem and bud culture. Plant tissue culture (PTC) is one among the group of biotechnological methods, utilizing in vitro strategies and techniques to culture living plant cells under controlled conditions. This technique primarily involves â ¦ The plant tissue culture industry has recorded a tremendous pace of growth reaching 500 million Mark in 1990 from a production of only 130 million in 1985-86. Plant tissue culture is a straightforward technique and many developing countries have already mastered it. Plant Tissue Culture is the process of growing isolated plant cells or organs in an artificial nutrient media outside the parent organism.. These topics are discussed from an academic as well industrial â ¦ Two different garlic ...
The experimental design required the analysis of four, partially confounded treatment groups; i.e. the TT treatment contains both B and M cell types found in TB and TM treatments, while the TM treatment combines the effects of cell-specificity and the stress effect that is also seen in TS. In the following discussion, the term stress is used to refer specifically to the effect of protoplast isolation on transcript levels. Importantly, although the aim of the experiment is to compare expression levels between B and M cell types, the level of gene expression within the intact M could never be directly measured because of the stress effect of M cell isolation. A model was constructed in order to formally describe this situation and to allow statistical elimination of the stress effect.. Let V 1f and V 2f represent the log2 transformed expression level of a given feature (f) in B and M cell types, respectively. The aim is to estimate (V 1f - V 2f ) on a feature-by-feature basis. We use c to label ...
Freshly isolated cell suspension protoplasts of Solanum dulcamara were mixed with Agrobacterium rhizogenes, allowed to settle for 2 h, exposed to electrical pulses and further incubated for 2h. Two...
BTC 501 Plant Biotechnology 3 CreditsThe plant biotechnology course covers principles and different aspects of plant biotechnology.Topics include:- Plant cell cultures; growing tissue-, axillary bud, root- and meristem cultures- Protoplast culture, somatic hybridisation and importance, cybridization- Application of mass propagation (micropropagation) for virus-free
Villanueva, M.A.; Metcalf, T.N.IIi; Wang, J.L., 1986: Monoclonal antibodies directed against protoplasts of soybean glycine max cells generation of hybridomas and characterization of a monoclonal antibody reactive with the cell surface
Jan. 15, 1974 H QGAWA ETAL 3,786,141 METHOD OF THE PRODUCTION OF ANTI-TUMOR SUBSTANCES Filed Sept. 50, 1971 United States Patent (3 3,786,141 METHOD OF PRODUCTION OF ANTI-TUMOR SUBSTANCES Haruki Ogawa, Fuchu, Akihiro Yamamoto, Higashimurayama, Yutaka Sugawara, Iruma-gun, and Shigeo Suzuki and Yoshio Takagalci, Kodaira, Japan, assignors to Chngai Seiyaku Kabushiki Kaisha, Tokyo, Japan Filed Sept. 30, 1971, Ser. No. 185,112 Claims priority, application Japan, Sept. 30, 1970, 45/ 85,006 Int. Cl. A01n /00 U.S. Cl. 424-95 10 Claims ABSTRACT OF THE DISCLOSURE A water-insoluble anti-tumor active substance is prepared from the living cells of hemolytic streptococci by lysing the cells of hemolytic streptococci with a lytic enzyme and collecting a water-insoluble fraction which corresponds to a protoplast membrane fraction of the cells. The preparation of the anti-tumor substance is far superior in the anti-tumor activity to that of water-soluble fraction and has less inflammation-, feverand paincausing ...
Transgenesis is the process of introducing an exogenous gene-called a transgene-into a living organism so that the organism will exhibit a new property and transmit that property to its offspring. Transgenesis can be facilitated by liposomes, enzymes, plasmid vectors, viral vectors, pronuclear injection, protoplast fusion, and ballistic DNA injection. Transgenesis can occur in nature. Transgenic organisms are able to express foreign genes because the genetic code is similar for all organisms. This means that a specific DNA sequence will code for the same protein in all organisms. Due to this similarity in protein sequence, scientists can cut DNA at these common protein points and add other genes. An example of this is the super mice of the 1980s. These mice were able to produce the human protein tPA to treat blood clots. The most common type of transgenesis research is done with bacteria and viruses which are able to replicate foreign DNA. The plasmid DNA is cut using restriction enzymes, ...
Zhicheng Zhang is the author of this article in the Journal of Visualized Experiments: mRNA Interactome Capture from Plant Protoplasts
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Martin Hughes (Bioc) (mjgh at uk.ac.cam.bio.mbfs) wrote: : ,In article ,26lcir$n6j at bigboote.WPI.EDU, eeyore at wpi.WPI.EDU ( eeYORE ) writes: : ,,Subject: viability assays : , : [about wanting a viability assay for roots : ,,I have tried FDA (fluorescein diacetate) stain, but this is not quantitative : ,,or reliable for plant tissue. Anyone have any ideas or experience? : ,, Melissa : I would also like to add a request. I have been using FDA to assay : for viability in protoplasts, which is a normal recomended method. : I am trying (as a control) to kill off protoplasts. Unfortunately, : follwing a death treatment (30 min in 0.1% NaAzide, or 0.3% KCyanide)- : which I imagine should be pretty fatal to the cells, the FDA stain : still shows positive (this is also the case with even higher toxin : levels). Any suggestions for a more sensitive method of viability : staining would be appreciated. I used neutral red which is accumulated in the vacuoles of living cells with a characteristic ...
CRISPRs capabilities have been expanded once again, this time to be used to increase the amount of healthy fats in soybeans. Lets skip right into the details for once without any backstory. To The Research! Researchers at the Institute for Basic Science (IBS) in South Korea have been working on their own variant of CRISPR […]. ...
Highly purified enzyme from Aspergillus niger used for isolation of biologically active protoplasts from a wide spectrum of higher plants and tissues.