Hygromycin b

Manufactured by Sangon

Sourced in China

Hygromycin B is a broad-spectrum antibiotic produced by the bacterium Streptomyces hygroscopicus. It functions as a protein synthesis inhibitor, preventing the translocation step during protein synthesis in prokaryotic and eukaryotic cells.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Hygromycin (7 citations)

11 protocols using hygromycin b

Lentiviral shRNA-Mediated Gene Depletion

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

To generate prostate cancer cells with stable depletion of indicated genes, lentiviral shRNA virus packaging and infection of cell lines were performed according to the protocol described previously [24 (link)]. In brief, lentiviral constructs were co-transfected with the pCMVdR8.91 (Delta 8.9) plasmid containing gag, pol, and rev genes, as well as the VSV-G envelope-expressing plasmid into HEK293T cells. Transfection with jetPRIME (Cat. No.101000046, Polyplus Transfection) was
performed based on the manufacturer’s instructions. The supernatant was harvested and filtered with a 0.45 mm syringe filter post-transfection at 48 h and 72 h. The medium containing virus was used for infecting indicated cell lines in the presence of Polybrene (4 mg/ml). Infected cells were selected using 2 µg/ml puromycin dihydrochloride hydrate (A610593-0025, Sangon Biotech) or 300 µg/ml hygromycin B (A100607-0100, Sangon Biotech) for 3 days.

Wu F., Wang M., Zhong T., Xiao C., Chen X., Huang Y., Wu M., Yu J, & Chen D. (2023). Inhibition of CDC20 potentiates anti-tumor immunity through facilitating GSDME-mediated pyroptosis in prostate cancer. Experimental Hematology & Oncology, 12, 67.

+ Open protocol

+ Expand

Characterization of Cell Lines for Immunoassay

Check if the same lab product or an alternative is used in the 5 most similar protocols

CHO-K1 and HEK-293 cells were obtained from American Type Culture Collection (ATCC CCL-61 and CRL-1573). HEK-293/NFκB-Luci/4-1BB cells were genetically engineered and expressed human 4-1BB and a luciferase reporter driven by a response element sensitive to 4-1BB agonistic stimulation and cultured in DMEM supplemented with 1 μg/mL puromycin (Gibco, C11995500BT) and 800 μg/mL hygromycin B (Sangon Biotech, A600230-0001). CHO-K1/CD32A, CHO-K1/CD32B, CHO-K1/CD16 and CHO-K1/hu4-1BB cells were designed to express the human Fcγ receptors (FcγRIIA, FcγRIIB, FcγIRA) and 4-1BB on the cell membrane, respectively, and grown in DMEM/F12 (HyClone, SH30023.01) containing 1 mg/mL Geneticin (Gibco, 11811023). The murine and human CRC cell lines CT26 and Colo205 were obtained from the cell bank affiliated with the Shanghai Institute of Biochemistry and Cell Biology (SIBCB), and the murine CRC cell line MC38 was purchased from Cobioer Company (Nanjing, China), authenticated, tested for mycoplasma contamination and cultured in RPMI-1640 medium (HyClone, SH30809.01). All media were supplemented with 10% fetal bovine serum (Ausbian, VS500T) and a 1% penicillin–streptomycin solution (HyClone, SV30010), and cells were cultured at 37 °C in a humidified incubator with 5% CO2.

Cheng L.S., Cheng Y.F., Liu W.T., Shen A., Zhang D., Xu T., Yin W., Cheng M., Ma X., Wang F., Zhao Q., Zeng X., Zhang Y, & Shen G. (2022). A humanized 4-1BB-targeting agonistic antibody exerts potent antitumor activity in colorectal cancer without systemic toxicity. Journal of Translational Medicine, 20, 415.

+ Open protocol

+ Expand

MMEJ-CRISPR Genome Editing in A. fumigatus

Check if the same lab product or an alternative is used in the 5 most similar protocols

For editing the A. fumigatus gene, the MMEJ-CRISPR system was used as described in our previous published papers [45 (link),49 (link)]. sgRNA targeted to the appropriate site of the target gene was synthesized in vitro using the MEGAscript T7 Kit (Life Technologies, AM1333). The corresponding repair template (DNA) with microhomology arms was amplified by PCR. Then, the repair template fragments and sgRNA were cotransformed into a Cas9-expressing A. fumigatus recipient strain. The primers and annotations for sgRNAs and repair templates are listed in Table S2. Transformation procedures were performed as previously described. Transformants were selected in medium lacking uridine or uracil or in the presence of 150 μg ml⁻¹ hygromycin B (Sangon) or 0.1 μg ml⁻¹ pyrithiamine (Sigma). For the recycling usage of the selectable marker pyr4, 1 mg ml⁻¹ 5-FOA was used for screening recipient strains. All primers used are listed in the supplementary data Table S2. All transformant isolates were verified by diagnostic PCR analysis using mycelia as the source of DNA. Primers were designed to probe upstream and downstream of the expected cleavage sites as labeled in Figure S1D.

Zhang C., Ren Y., Gu H., Gao L., Zhang Y, & Lu L. (2021). Calcineurin-mediated intracellular organelle calcium homeostasis is required for the survival of fungal pathogens upon extracellular calcium stimuli. Virulence, 12(1), 1091-1110.

+ Open protocol

+ Expand

Fungal Growth under Iron and Calcium Conditions

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The strains used in this study are summarized in Table S1. All chemicals used in this study were analytic reagents from the indicated companies or from Sigma-Aldrich Co. Generally, A. fumigatus strains were grown on minimal medium (MM) containing 1% (wt/vol) glucose and 70 mM NaNO₃ as the sole carbon and nitrogen sources, respectively. MM+UU contained MM plus 5 mM uridine and 10 mM uracil. For iron-deficient conditions, iron was omitted from the trace element solution; for harsh iron-deficient conditions, the iron-specific chelator bathophenanthroline disulfonate (BPS) was added to the iron-deficient medium. Supplementation with calcium was carried out as described in the figure legends. Transformants were screened on medium containing 200 μg/mL hygromycin B (Sangon Biotech; A600230). The strains Cryptococcus neoformans H99 and Candida albicans ATCC 10231 were used for the experiments and maintained on yeast nitrogen base (YNB) medium (0.67% YNB medium, 2% dextrose) (74 (link)). Growth under low-iron conditions was performed in YNB with the addition of BPS (YNB-BPS). Supplementation with calcium was carried out or omitted, depending on the circumstances.

Ye J., Wang Y., Li X., Wan Q., Zhang Y, & Lu L. (2022). Synergistic Antifungal Effect of a Combination of Iron Deficiency and Calcium Supplementation. Microbiology Spectrum, 10(3), e01121-22.

+ Open protocol

+ Expand

Fungal Growth under Iron and Calcium Conditions

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ye J., Wang Y., Li X., Wan Q., Zhang Y, & Lu L. (2022). Synergistic Antifungal Effect of a Combination of Iron Deficiency and Calcium Supplementation. Microbiology Spectrum, 10(3), e01121-22.

+ Open protocol

+ Expand

Fungal Strain Cultivation and Transformation

Check if the same lab product or an alternative is used in the 5 most similar protocols

All A. fumigatus strains used in this study are summarized in Table 1. In general, the strains were grown on a minimal medium (MM) or MMUU (MM supplemented with 5 mM uridine and 10 mM uracil) pH 6.5, containing 50 mL/L salt, 1% glucose, 1 mL/L 1,000 × trace elements, and 2% agar (Gupta et al., 1976 (link)). For all liquid media, agars were spared. The transformation was performed as per the protocol (Szewczyk et al., 2006 (link)), and hygromycin B (200 μg/mL, Shanghai Sangon Co., China) was used as a selection marker. All strains were cultured at 37°C for 1.5–3 days.

Zhou X., Yang G., Li C., Yang F, & Chang X. (2022). Requirement of a putative mitochondrial GTPase, GemA, for azole susceptibility, virulence, and cell wall integrity in Aspergillus fumigatus. Frontiers in Microbiology, 13, 957857.

+ Open protocol

+ Expand

Culturing and Characterizing A. fumigatus

Check if the same lab product or an alternative is used in the 5 most similar protocols

A. fumigatus strains, used in this study, are summarized in S2 Table. Generally, A. fumigatus strains were grown on minimal medium (MM)[33 (link)] containing 1% (w/v) glucose and 70 mM NaNO₃ as sole carbon and nitrogen sources, respectively. For iron starvation, iron was omitted in the trace element solution; for increased iron starvation, the iron-specific chelator bathophenanthroline disulfonate (BPS) was added in iron depleted media. Supplementation with iron (FeCl₃) and/or leucine was carried out as described in the Figures. Transformants were screened on media containing 200 μg/ml hygromycin B (Shanghai Sangon Co., China). To analyze the phenotype of the mutants, 2 x 10³ conidia were point-inoculated on plates. All plates were incubated at 37°C for two days.

Long N., Orasch T., Zhang S., Gao L., Xu X., Hortschansky P., Ye J., Zhang F., Xu K., Gsaller F., Straßburger M., Binder U., Heinekamp T., Brakhage A.A., Haas H, & Lu L. (2018). The Zn2Cys6-type transcription factor LeuB cross-links regulation of leucine biosynthesis and iron acquisition in Aspergillus fumigatus. PLoS Genetics, 14(10), e1007762.

+ Open protocol

+ Expand

Genetic Modifications in Aspergillus

Check if the same lab product or an alternative is used in the 5 most similar protocols

Strains (ΔpyrG and brlA^xylP) were constructed by the CRISPR-Cas9 system as described previously [22 (link),24 (link)]. For the deletion of the pyrG gene (ASPNIDRAFT_128428), a co-transformation was performed using pFC332 and an in vitro synthesized sgRNA targeting pyrG. This was achieved through PEG4000-mediated protoplast transformation [24 (link)], with the added selection pressures of 5-Fluoroorotic acid (5-FOA, Sangon Biotech, SA601555, Nanjing, China) and hygromycin B (Sangon Biotech, A600230, Nanjing, China) resulting in the creation of the ΔpyrG strain. The final strain, brlA^xylP, was obtained by removing pFC330 under the stress of 5-FOA and introducing a pyrG fragment for complementation. To ensure the accuracy of the genetic modifications, all strains underwent verification through sequencing and/or diagnostic PCR targeting the relevant editing genes.

Wu X., Zhang T., Zhang K., Zhang R., Shi M., Gu C., Shi T., Lu L., Xue F., Xu Q, & Zhang C. (2024). The forced activation of asexual conidiation in Aspergillus niger simplifies bioproduction. Synthetic and Systems Biotechnology, 9(2), 277-284.

+ Open protocol

+ Expand

Synthetic Lethality Screening of Kinetochore Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Standard media [either YE (yeast extract) rich medium or EMM (Edinburgh minimal medium)] and culturing methods were used (Moreno et al. 1991 (link); Forsburg and Rhind 2006 (link)). G418 disulfate (Sigma-Aldrich), hygromycin B (Sangon Biotech), or nourseothiricin (clonNAT; Werner BioAgents) was used at a final concentration of 100 μg/ml and thiabendazole (TBZ) (Sigma-Aldrich) at 5–15 μg/ml in YE media. For serial dilution spot assays, 10-fold dilutions of a mid-log-phase culture were plated on the indicated media and grown for 3–5 days at indicated temperatures. To examine the possible synthetic lethality (SL) of genetic combinations between alleles of GBP-mCherry, GFP-tagged kinetochore proteins, and spindle checkpoint mutants mad2Δ or bub1Δ, normal-looking 4-spore asci obtained after crosses between parental strains were dissected using a micromanipulator. At least 20 complete tetrads were dissected after each genetic cross, and the genotypes of colonies formed from germinated spores were deduced after being replicated on selective plates. The frequency of spores with expected genotypes failing to germinate was quantified, it was classified as SL, strong growth defect, or normal growth when the frequency was above 80%, between 20% and 80%, or below 20%, respectively. Yeast strains used and created in this study are listed in Supplementary Table S1.

Deng D.J., Xia Q.C., Jia G.S., Suo F., Chen J.L., Sun L., Wang J.Q., Wang S.M., Du L.L., Wang Y, & Jin Q.W. (2021). Perturbation of kinetochore function using GFP-binding protein in fission yeast. G3: Genes|Genomes|Genetics, 11(11), jkab290.

+ Open protocol

+ Expand

Extraction and Characterization of Bioactive Compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols

Methyl jasmonate (95%), apigenin 7-O-glucuronide, scutellarin, apigenin, scutellarein, apigenin 7-O-glucoside, naringenin chalcone, naringenin (98%), and acetosyringone were all purchased from Sigma (United States). The antibiotics cefotaxime and hygromycin B were purchased from Sangon (China). The plant hormones 1-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (6-BA) were purchased from Phytechnology Laboratories (United States). Ammonium acetate was purchased from Thermo Fisher Scientific (United States) and methanoic acid from Aladdin (China). Methanol and acetonitrile of high performance liquid chromatography (HPLC) grade were from Merck Company (Germany). A Milli-Q Reagent Water System was used to produce ultrapure water (Millipore, United States).

Chen R., Chen X., Zhu T., Liu J., Xiang X., Yu J., Tan H., Gao S., Li Q., Fang Y., Chen W., Zhang L, & Huang B. (2018). Integrated Transcript and Metabolite Profiles Reveal That EbCHI Plays an Important Role in Scutellarin Accumulation in Erigeron breviscapus Hairy Roots. Frontiers in Plant Science, 9, 789.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!