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Canavanine

Q: Are there different types or variations of Canavanine?

While Canavanine is a single, distinct non-proteinogenic amino acid, there may be slight variations in its chemical structure or properties depending on the plant source or extraction method. Researchers should be aware of these potential differences and ensure they are using the appropriate form of Canavanine for their specific study.

Canavanine is a non-proteinogenic amino acid found in various plant species, particularly in legumes.
It is structurally similar to the essential amino acid arginine and can interfere with its biological functions.
Canavanine has been studied for its potential therapeutic applications, such as in cancer treatment and as a defense mechanism in plants against herbivores.
Researchers can leverage PubCompare.ai's AI-driven platform to streamline their Canavanine research, easily locating relevant protocols and optimizing their experimental approach to drive reproducible results.

Most cited protocols related to «Canavanine»

Yeast Genetic Manipulation Using Plasmids and Selective Media

Cited 32 times

Propagation of plasmids was performed in chemically competent Escherichia coli DH5α according to manufacturer instructions (Z-competent™ transformation kit; Zymo Research, CA). All yeast strains used in this study are listed in Table 2. Under nonselective conditions, yeast was grown in complex medium (YPD) containing 10 g L⁻¹ yeast extract, 20 g L⁻¹ peptone, and 20 g L⁻¹ glucose. Synthetic media (SM) containing 3 g L⁻¹ KH₂PO₄, 0.5 g L⁻¹ MgSO₄·7H₂O, 5 g L⁻¹ (NH₄)₂SO₄, 1 mL L⁻¹ of a trace element solution as previously described (Verduyn et al., 1992 (link)), 1 mL L⁻¹ of a vitamin solution (Verduyn et al., 1992 (link)) were used. When amdSYM was used as marker, (NH₄)₂SO₄ was replaced by 0.6 g L⁻¹ acetamide as nitrogen source and 6.6 g L⁻¹ K₂SO₄ to compensate for sulfate (SM-Ac). Recycled markerless cells were selected on SM containing 2.3 g L⁻¹ fluoroacetamide (SM-Fac). SM, SM-Ac, and SM-Fac were supplemented with 20 mg L⁻¹ adenine and 15 mg L⁻¹l-canavanine sulfate when required. In all experiments, 20 g L⁻¹ of glucose was used as carbon source. The pH in all the media was adjusted to 6.0 with KOH. Solid media were prepared by adding 2% agar to the media described above.

Solis-Escalante D., Kuijpers N.G., Bongaerts N., Bolat I., Bosman L., Pronk J.T., Daran J.M, & Daran-Lapujade P. (2012). amdSYM, a new dominant recyclable marker cassette for Saccharomyces cerevisiae. Fems Yeast Research, 13(1), 126-139.

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Publication 2012

acetamide Adenine Agar Canavanine Carbon Cells Escherichia coli fluoroacetamide Glucose Nitrogen Peptones Plasmids Strains Sulfate, Magnesium Sulfates, Inorganic Trace Elements Vitamin A Yeast, Dried

Cryptococcosis Isolate Identification and Preservation

Cited 23 times

Cryptococcal isolates studied in this study are listed in Appendix Tables 1 and 2. The isolates were obtained by the participating laboratories of the IberoAmerican Cryptococcal Study Group and maintained on Sabouraud dextrose agar slants at 4°C and as water cultures at room temperature. Isolates were identified as C. neoformans by using standard methods (20 ). The variety was determined by the color reaction test on L-canavanine-glycine-bromothymol blue medium (21 (link)), and the serotype was determined, in selected isolates, by the use of the Crypto Check Kit (Iatron Laboratories Inc., Tokyo, Japan).
The isolates were sent for molecular typing to the Molecular Mycology Laboratory at the University of Sydney at Westmead Hospital, Sydney, Australia, either as water cultures or on Sabouraud dextrose agar slants. For long-term storage, the isolates were maintained as glycerol stocks at –70°C.

Meyer W., Castañeda A., Jackson S., Huynh M, & Castañeda E. (2003). Molecular Typing of IberoAmerican Cryptococcus neoformans Isolates. Emerging Infectious Diseases, 9(2), 189-195.

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Publication 2003

Agar Bromthymol Blue Canavanine Cryptococcus Cryptococcus neoformans Glucose Glycerin Glycine

Yeast Genetic Manipulation and Microscopy

Cited 12 times

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Publication 2011

Alleles Biological Assay Canavanine derivatives Diet, Protein-Restricted DNA Replication Gene Deletion Genes, Reporter Glucose Green Fluorescent Proteins HSP40 Heat-Shock Proteins Microscopy Mismatch Repair MSH6 protein, human Mutation Peptones Plasmids Point Mutation Protein C Proteins Recombination, Genetic red fluorescent protein Saccharomyces cerevisiae Strains

Yeast Genetic Modification Protocol

Cited 10 times

Yeast transformations were performed using the lithium acetate protocol (Gietz & Woods, 2002 (link)). Integration of the deletion cassettes into the yeast genome was selected by plating the transformation mix on SM-Ac. Targeted integration was verified by PCR with the primer pairs CdcFW/CdcRV, AdcFW/AcdRV, HdcFW/HdcRV, ScARO80dcFW/amdSdcRV, ScARO80dcFW/ScARO80gRV, SbARO80dcFW/amdSdcRV, SbARO80dcFW/SbARO80gRV and, when applicable, by transferring single colonies to SM plates containing 15 mg L⁻¹l-canavanine or by screening for colony pigmentation. A small fraction of single colony was resuspended in 15 μL of 0.02 N NaOH; 2 μL of this cell suspension was used as template for the PCR that was performed using DreamTaq PCR master mix (Fermentas GmbH, St. Leon-Rot, Germany) following the manufacturer recommendations. Marker removal was achieved by growing cells overnight in liquid YPD and transferring 0.2 mL to a shake flask containing 100 mL of SM-FAc. Marker-free single colonies were obtained by plating 0.1 mL of culture on SM-Fac solid media and confirmed by PCR with the primer pairs CdcFW/CdcRV or AdcFW/AcdFW. All cultures were incubated at 30 °C. To confirm that only endogenous sequences were present after marker removal, long run sequencing (Baseclear, Leiden, the Netherlands) was performed using the primers C-FW and C-RV for the CAN1 locus and A-FW and A-RV for ADE2 locus of the marker-free strain IMX206.

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Publication 2012

Canavanine Cells Deletion Mutation Genome lithium acetate Oligonucleotide Primers Pigmentation Strains Tremor Yeast, Dried

Analyzing Spontaneous and Induced Crossovers in Yeast

Cited 7 times

All experiments were conducted with the diploid strain PG311 (Lee et al. 2009 (link)). The relevant genotype of PG311 is MATα::NAT^R/MATaURA3/ura3-1 ade2-1/ade2-1 TRP1/trp1-1 HIS3/his3-11,15 GAL2/gal2SUP4-o/can1-100 V9229/V9229::HYG V261553/V261553::LEU2. This diploid was generated by crossing the haploid strains PSL2 and PSL5, which are isogenic with strains W303a and YJM789, respectively, except for alterations introduced by transformation (Lee et al. 2009 (link)). From this point on, we will refer to the haploid parents of PG311 as W303a and YJM789. In general, PG311 has the SNPs predicted from the haploid parents. The disruption of MATα in PG311 allows synchronization of this diploid by α-factor. Although diploids that lack MATα do not sporulate under normal conditions, such strains can be sporulated on plates containing 5 mM nicotinamide (J. Rine, personal communication). For experiments in which we analyzed meiotic products of PG311, the stains were pregrown on YPD plates with 5 mM of nicotinamide and then transferred to sporulation plates containing 5 mM nicotinamide. Plates were incubated at 25° for 2–4 days before tetrad dissection.
Standard media were used (Guthrie and Fink 1991 ) unless noted. To detect spontaneous crossovers, we first isolated single colonies of PG311 grown on rich growth medium (YPD) at 30° for 2 days. Individual colonies were suspended in 400 μl of dH₂O, and 100 μl of this mixture was plated on canavanine-containing plates (SD-arg + 120 μg/ml canavanine). The plates were incubated 4 days at room temperature, followed by incubation for 16 hr at 4°; the 4° incubation allows better visualization of the red sectors. We purified cells from the red and white sectors for subsequent analysis.
In the experiments in which cells were irradiated, we synchronized cells in G1 using α-factor (Lee and Petes 2010 (link)). The synchronized cells were treated with either γ-radiation in a Shepherd Mark 1 ¹³⁷Cs irradiator at 100 Gy or with UV (254 nm) derived from a TL-2000 Ultraviolet Translinker at a dosage of 10 or 15 J/m². Following radiation, the cells were plated either on nonselective plates (SD-arg) or plates that lacked arginine and contained 120 μg/ml canavanine. The subsequent growth of the cells and the analysis of sectors were done as described above for the spontaneous selection with the exception that sectored colonies for the UV-treated samples were isolated from nonselective plates grown at 30° instead of room temperature.

St. Charles J., Hazkani-Covo E., Yin Y., Andersen S.L., Dietrich F.S., Greenwell P.W., Malc E., Mieczkowski P, & Petes T.D. (2012). High-Resolution Genome-Wide Analysis of Irradiated (UV and γ-Rays) Diploid Yeast Cells Reveals a High Frequency of Genomic Loss of Heterozygosity (LOH) Events. Genetics, 190(4), 1267-1284.

Publication 2012

Arginine Canavanine Culture Media Diploidy Dissection Erythrocytes Genotype Miotics Niacinamide Parent Radiation Single Nucleotide Polymorphism Staining Strains tyrosinase-related protein-1

Most recents protocols related to «Canavanine»

Quantifying CAN1 Mutation Rates

The CAN1 fluctuation analysis was performed as described in^{57 (link)}. Relevant genotypes for the Can^R mutation assay were streaked out 48 h prior inoculation to conserve population doublings within replicates. At least 14 independent single colonies from each genotype were entirely excised from the agar plate using a sterile scalpel to inoculate a 10 ml of YPD medium. The cultures were incubated at 30 °C, 250 rpm for 16 h. After measuring the optical density of the cultures they were harvested by centrifugation. Then, each culture was resuspended in 1 ml sterile water. Exactly 1 ml of each resuspension was transferred to a new tube. From this, a 10-fold dilution series up to a dilution factor of 10⁶ was performed in a 96-well plate. Finally, 100 µl of all strains from the 10⁻⁶ dilution were plated on a YPD plate and distributed with exactly four glass beads per plate. All strains were plated on SC-ARG plates supplemented with 60 µg/mL canavanine with the indicated dilution factor. The plates were incubated for 72 h at 30 °C before the outgrown colonies were manually counted. The medium for the Can^R mutation assay was mixed, autoclaved and poured each day before plating to maintain constant conditions between replicates.
For evaluation, the number (#) of mutant cells per culture, representing r, was calculated:

r = c u l t u r e v o l . \times (c o n c . f a c t o r \times \frac{# m u t a n t c o l o n i e s}{p l a t e d v o l .} \times d i l u t i o n f a c t o r (p l a t e d))

The following correction was used to account for the progenies of each individual CAN1 mutation event per cell. With M being a scaled value that represents the number of cells that have actually undergone a mutation event (from which the counted progenies originated):

r = M (1.24 + \ln M)

The final mutation rate was calculated dividing M by the total number of cells present in the initial culture:

m u t a t i o n r a t e = \frac{M}{# c e l l s i n t h e c u l t u r e}

The data was plotted as the Median with 95% Confidence interval using the GraphPad PRISM8 software.

Schindler N., Tonn M., Kellner V., Fung J.J., Lockhart A., Vydzhak O., Juretschke T., Möckel S., Beli P., Khmelinskii A, & Luke B. (2023). Genetic requirements for repair of lesions caused by single genomic ribonucleotides in S phase. Nature Communications, 14, 1227.

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Publication 2023

Agar Biological Assay Canavanine Cell Culture Techniques Cells Centrifugation Genotype M Cells Mutation R Factors Sterility, Reproductive Strains Technique, Dilution Vaccination Vision

Yeast Strain Construction for SGA Analysis

The G1-RNH202-TAP (this study), S-, and G2-RNH202-TAP alleles^{8 (link)} were crossed to the haploid background strain (Y8205, Source C. Boone) for the SGA query strain construction. Selection of diploids, sporulation and tetrad analysis generated the four query strains used in SGA analysis. Cell cycle restricted protein expression of Rnh202 was confirmed by arrest and release experiment and western blot analysis. The selectable markers for SGA analysis were verified by PCR (oMT86/oMT91 for can1Δ::STE2pr-Sp_his5, oMT89/oMT90 for lyp1Δ::STE3pr-LEU2) and replica-plating on YPD + (50 μg/ml canavanine, 50 μg/ml thialysine). The yeast strains are listed in Supplementary Data 2.

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Publication 2023

Canavanine Cell Cycle Diploidy Proteins S-2-aminoethyl cysteine Saccharomyces cerevisiae Strains Western Blot

Detecting Mutations and Recombination in Yeast

Derivatives of E134 were plated on media that allowed us to detect new mutations: CAN medium is a standard minimal medium that lacks arginine and contains canavanine, a toxic agent that is an analog to arginine. If loss of function mutations occur in the CAN1 gene, the transporter loses its function, preventing the toxic canavanine from entering the cell and allowing the mutated cells to grow on this medium. The LYS2 locus of E134-derived strains contains an insertion of 14 adenine residues, which causes auxotrophy to lysine. Frameshift mutations (+1 or −2) can restore the Lys+ phenotype. SD-Lys is a standard-defined medium that lacks lysine. The trp1-289 mutation of E134-derived strains contains a single nucleotide change, which causes auxotrophy to tryptophan. Reversion to the original nucleotide allows the cells to grow on an SD-Trp plate, which is a standard-defined medium that lacks tryptophan.
Derivatives of BLS2 were plated on media that allowed us to detect new sister chromatid recombination: Strain BLS2 contains 5′ ade3 and 3′ ade3 fragments that are separated by URA3 marker. This construct allows the detection of new unequal sister chromatid recombination (USCR) by plating the cells on the appropriate medium (SD-HIS) [32 (link)]. Mutation and USCR rates were calculated as described in [33 (link)].

Nir Heyman S., Golan M., Liefshitz B, & Kupiec M. (2023). A Role for the Interactions between Polδ and PCNA Revealed by Analysis of pol3-01 Yeast Mutants. Genes, 14(2), 391.

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Publication 2023

Adenine Arginine Canavanine Chromatids derivatives Frameshift Mutation Genes Loss of Function Mutation Lysine Membrane Transport Proteins Mutation Nucleotides Phenotype Recombination, Genetic Strains Tryptophan tyrosinase-related protein-1

Measuring UV-Induced Mutagenesis via CAN1 Assay

UV-induced mutagenesis was measured with the CAN1 forward-mutation assay [59 (link)]. UV-induced mutagenesis frequencies were obtained by dividing the number of colonies growing on synthetic plates without arginine and containing l-canavanine (Sigma, 30 mg/l) (i.e., canavanine-resistant, Can^R, cells) by the number of cells that survived irradiation counted on rich media YPD. The number of Can^R colonies obtained after irradiation was corrected by subtracting the number of spontaneous Can^R colonies growing on non-irradiated plates.

Maloisel L., Ma E., Phipps J., Deshayes A., Mattarocci S., Marcand S., Dubrana K, & Coïc E. (2023). Rad51 filaments assembled in the absence of the complex formed by the Rad51 paralogs Rad55 and Rad57 are outcompeted by translesion DNA polymerases on UV-induced ssDNA gaps. PLOS Genetics, 19(2), e1010639.

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Publication 2023

Arginine Biological Assay Canavanine Cells Mutagenesis Mutation

Genetic Screening for Mating and Meiosis Defects

A centromere-based plasmid carrying URA3 and KAR4 with an internal 3xHA tag under the control of its native promoter (pMR2654) was mutagenized with hydroxylamine (see Table S2). The plasmid (10 μg) was incubated with 500 μl of 7% hydroxylamine solution for 20 hours at 37° (Rose, Winston et al. 1990 ). The reaction was stopped by adding 10 μl 5M NaCl and 50 μl 1mg/ml BSA, and the DNA was ethanol-precipitated and resuspended in TE. The mutagenized DNA was transformed into the yeast strain MY 10128 (see Table S1). Individual transformants were patched onto SC-URA media, 50 to a plate, with a positive and negative control (MY 11596 and 11597). The patches were replica-printed onto YEPD with a lawn of MATα MY11297 and allowed to mate at 30° C for 3 hours (limited mating) before replica-printing on SC-LEU-LYS media to detect mating defects. The mating plates were returned to the 30° C incubator overnight, then replica printed onto SC-LEU-LYS-URA media the next day to select for diploids from all patches (overnight mating). Diploids from the overnight mating were replica printed onto Complete Sporulation media and incubated at room temperature for one day, then at 30° for three days. The sporulation plates were finally replica printed on SC-HIS-ARG media with added canavanine to detect successful entry into meiosis. The limited mating plates were compared to the meiosis plates to detect potential separation of function mutants. Potential hits were repatched (nine to a plate) from the original transformation plate and rescreened. When transformants displayed a consistent phenotype, the plasmid was extracted using a yeast mini-prep kit (Zymo Research, Irvine, CA) and electroporated into bacteria. Bacterial colonies were grown up and the plasmids were extracted using a kit (Qiagen, Germantown, MD). The samples were sent for sequencing with KAR4-specific primers. After sequencing, the plasmids were retransformed into MY 10128 and assayed a final time for both mating and meiotic defects. To further address the severity of the meiotic defect, cells were sporulated as described above and spotted in 10-fold serial dilutions on the SC-HIS-ARG media. Alleles that supported wild type levels (formed colonies at the highest dilution in the series) of recombination were scored as “++++”. Alleles that only formed colonies on the 10⁻² serial dilution were scored as “+++” and so on.

Park Z.M., Sporer A., Kraft K., Lum K., Blackman E., Belnap E., Yellman C, & Rose M.D. (2023). Kar4, the Yeast Homolog of METTL14, is Required for mRNA m6A Methylation and Meiosis. bioRxiv.

Publication Preprint 2023

Alleles Bacteria Canavanine Cells Centromere Diploidy Ethanol Hydroxylamine leucyl-lysine Meiosis Miotics Oligonucleotide Primers Phenotype Plasmids Printed Media Recombination, Genetic Saccharomyces cerevisiae Sodium Chloride Strains Technique, Dilution

Top products related to «Canavanine»

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Canavanine is a laboratory reagent produced by Merck Group. It is a non-proteinogenic amino acid that can be used as a biochemical tool in research applications.

L canavanine by Merck Group

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L-canavanine is a laboratory reagent that serves as a structural analog of the amino acid L-arginine. It is commonly used in biochemical and cell biology research to study the effects of arginine analogues on various cellular processes and pathways.

Thialysine by Merck Group

Sourced in Germany

Thialysine is a laboratory product manufactured by Merck Group. It is a chemical compound used in various research and analytical applications. The core function of Thialysine is to serve as a chemical reagent, without providing any further interpretation or extrapolation on its intended use.

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What is Canavanine?

How can PubCompare.ai help with Canavanine research?

PubCompare.ai allows you to screen protocol literature more efficiently and leverage AI to pinpoint critical insights. The platform can help researchers identify the most effective protocols related to Canavanine for their specific research goals. PubCompare.ai's AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy.

What are some common challenges with using Canavanine?

One of the main challenges with using Canavanine is its structural similarity to arginine, which can lead to interference with arginine's biological functions. Researchers must be careful to account for this when designing experiments and interpreting results. Additionally, the availability and purity of Canavanine samples can vary, which can impact the consistency and reliability of research findings.

Are there different types or variations of Canavanine?

What are some of the key applications of Canavanine?

Canavanine has been studied for its potential therapeutic applications, such as in cancer treatment and as a plant defense mechanism against herbivores. Researchers are also investigating its use as a tool for studying arginine-dependent biological processes and pathways. Additionally, Canavanine may have potential applications in the development of novel pharmaceuticals or agrochemicals, though more research is needed to fully explore its capabilities.

More about "Canavanine"

Canavanine, a non-proteinogenic amino acid, is a naturally occurring compound found in various plant species, particularly legumes.
It is structurally similar to the essential amino acid arginine and can interfere with its biological functions.
This unique compound has garnered significant interest from researchers due to its potential therapeutic applications, such as in cancer treatment, and its role as a defense mechanism in plants against herbivores.
Canavanine, also known as L-canavanine, is a close relative of the amino acid thialysine, another non-proteinogenic compound.
Researchers can leverage advanced tools like the RoToR bench-top colony arrayer to study the effects of canavanine on various organisms, including microbes.
These experiments may involve the use of other compounds like hygromycin B, a selective antibiotic, and yeast nitrogen base, a medium for culturing yeast.
Beyond its biological applications, canavanine has also been studied for its potential use in food and agriculture.
For instance, monosodium glutamate (MSG), a common food additive, has been examined for its interaction with canavanine.
Additionally, ampicillin, a widely used antibiotic, has been explored in the context of canavanine research.
To facilitate their canavanine studies, researchers can utilize powerful software like Quantity One, which provides advanced analysis and visualization tools.
By leveraging these resources, scientists can streamline their experimental processes and gain valuable insights into the diverse applications of this unique amino acid.
Whether exploring canavanine's therapeutic potential, its role in plant-herbivore interactions, or its broader implications in food and agriculture, researchers can trust PubCompare.ai's AI-driven platform to optimize their research workflows and drive reproducible results.
With access to relevant protocols, pre-prints, and patents, researchers can navigate the canavanine landscape with confidence and efficiency, ultimately advancing our understanding of this fascinating compound.