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Hygromycin B

(Synonyms: 潮霉素; 潮霉素B; Hygrovetine) 目录号 : GC15496

Hygromycin B 是一种氨基糖苷类抗生素,对原核和真核细胞具有活性。

规格 价格 库存 购买数量
10mM (in 1mL Water)
¥200.00
现货
500mg
¥378.00
现货
1g
¥620.00
现货

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Sample solution is provided at 25 µL, 10mM.

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实验参考方法

Cell experiment [1]:

Cell lines

Wild type haploid Saccharomyces cerevisiae strain Y166 and the spontaneous CRY6 mutant

Preparation Method

Logarithmically growing yeast cells (about 2- 107/ml) were converted into spheroplasts by treatment with glusulase. The spheroplasts were recovered by incubation at 30℃ for 90 min in YEPD medium plus I M sorbitol. Then the culture was divided in three aliquots and each received 10 µCi/ml of [3H]leucine (54 Ci/mmol). One aliquot served as control, the other two received 250 and 500 µg/ml hygromycin B, respectively, and incubation was continued at 30℃.

Reaction Conditions

50 and 500 µg/ml,30℃,20-120 min.

Applications

Protein synthesis by yeast spheroplasts is blocked by hygromycin B as determined by the uptake of [3H]leucine into trichloroacetic acid-precipitable material

References:

[1]: Gonzalez A, Jimenez A, Vazquez D, Davies JE, Schindler D. Studies on the mode of action of hygromycin B, an inhibitor of translocation in eukaryotes. Biochim Biophys Acta 1978; 521:459-469.

产品描述

Hygromycin B is an aminoglycoside antibiotic produced by Streptomyces hygroscopicus. Widely used in veterinary medicine and in cell culture selections, it kills bacteria, fungi, and higher eukaryotic cells, including mammalian cells [1].

The sensitivity of various cultured cell lines to Hygromycin B was assessed by plating cells at low density in Dulbecco modified Eagle medium supplemented with 10% fetal calf serum and Hygromycin B to give final drug concentrations ranging between 50 and 400 ug/ml of medium. Usually one or two cycles of replication still occurred before the onset of cytotoxicity; cell death commenced ca.3 days after the beginning of drug treatment and was generally complete after 8days (CV1 and HeLa cells sometimes required 10 to 12 days before cell killing was complete). Sofar, no cellline has been found that is naturally resistant to Hygromycin B [2].

The E.coli bacterial Hygromycin B resistance gene provides a basis for testing the usefulness of Hygromycin B as adominant selectable marker in Hygromycin B -susceptible cells. It may also be possible to use promoter less coding sequences of the Hygromycin B resistance gene as a promoter probe [3].

Hygromycin B provides a generally applicable selection system for DNA transfer experiments between both procaryotic and eukaryotic cells [2].

References:
[1]. Borovinskaya MA, Shoji S, Fredrick K, Cate JH. 2008. Structural basis for hygromycin B inhibition of protein biosynthesis. RNA 14: 1590–99
[2]. KAREN BLOCHLINGER,HEIDI DIGGELMANN. Hygromycin B Phosphotransferase as a Selectable Markerfor DNA Transfer Experiments with Higher Eucaryotic Cells. MOLECULAR AND CELLULAR BIOLOGY, Dec.1984, p.2929-2931
[3]. R. N. Rao, N. E. Allen, J. N. J. Hobbs, W. E. J. Alborn, H. A. Kirst & J. W. Paschal: Genetic and enzymatic basis of hygromycin B resistance in Escherichia coli. Antimicrob. Agents Chemother. 24, 689-695 (1983)

Chemical Properties

Cas No. 31282-04-9 SDF
别名 潮霉素; 潮霉素B; Hygrovetine
化学名 (3'R,3aS,4S,4'R,5'R,6R,6'R,7S,7aS)-4-[(1R,2S,3R,5S,6R)-3-amino-2,6-dihydroxy-5-(methylamino)cyclohexyl]oxy-6'-[(1S)-1-amino-2-hydroxyethyl]-6-(hydroxymethyl)spiro[4,6,7,7a-tetrahydro-3aH-[1,3]dioxolo[4,5-c]pyran-2,2'-oxane]-3',4',5',7-tetrol
Canonical SMILES CNC1CC(C(C(C1O)OC2C3C(C(C(O2)CO)O)OC4(O3)C(C(C(C(O4)C(CO)N)O)O)O)O)N
分子式 C20H37N3O13 分子量 527.5
溶解度 ≥ 26.375mg/mL in Water 储存条件 Store at -20°C
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

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Research Update

Exploration of Hygromycin B Biosynthesis Utilizing CRISPR-Cas9-Associated Base Editing

ACS Chem Biol2020 Jun 19;15(6):1417-1423.PMID: 32275383DOI: 10.1021/acschembio.0c00071

Hygromycin B is an aminoglycoside antibiotic widely used in industry and biological research. However, most of its biosynthetic pathway has not been completely identified due to the immense difficulty in genetic manipulation of the producing strain. To address this problem, we developed an efficient system that combines clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9-associated base editing and site-specific recombination instead of conventional double-crossover-based homologous recombination. This strategy was successfully applied to the in vivo inactivation of five candidate genes involved in the biosynthesis of hygromycin B by generating stop codons or mutating conserved residues within the encoding region. The results revealed that HygJ, HygL, and HygD are responsible for successive dehydrogenation, transamination, and transglycosylation of nucleoside diphosphate (NDP)-heptose. Notably, HygY acts as an unusual radical S-adenosylmethionine (SAM)-dependent epimerase for hydroxyl carbons, and HygM serves as a versatile methyltransferase in multiple parallel metabolic networks. Based on in vivo and in vitro evidence, the biosynthetic pathway for hygromycin B is proposed.

Enhanced resistance of Trichoderma harzianum LZDX-32-08 to hygromycin B induced by sea salt

Biotechnol Lett2021 Jan;43(1):213-222.PMID: 32851464DOI: 10.1007/s10529-020-02994-y

Objectives: To determine the effect of sea salt on the resistance of Trichoderma harzianum LZDX-32-08 to hygromycin B and speculate the possible mechanisms involved via transcriptome analysis.
Results: Sea salt addition in media to simulate marine environment significantly increased the tolerance of marine-derived fungus Trichoderma harzianum LZDX-32-08 to hygromycin B from 40 to 500 ¦̧/ml. Meanwhile, sea salt addition also elicited the hygromycin B resistance of 5 other marine or terrestrial fungi. Transcriptomic analyses of T. harzianum cultivated on PDA, PDA supplemented with sea salt and PDA with both sea salt and hygromycin B revealed that genes coding for P-type ATPases, multidrug resistance related transporters and acetyltransferases were up-regulated, while genes coding for Ca2+/H+ antiporter and 1,3-glucosidase were down-regulated, indicating probable increased efflux and inactivation of hygromycin B as well as enhanced biofilm formation, which could jointly contribute to the drug resistance.
Conclusions: Marine environment or high ion concentration in the environment could be an importance inducer for antifungal resistance. Possible mechanisms and related key genes were proposed for understanding the molecular basis and overcoming this resistance.