Biosynthesis of Hygromycin-B Antibiotic by Streptomyces crystallinus AZ151 Isolated from Assuit, Egypt

2012 ◽  
Vol 2 (3) ◽  
pp. 46-65 ◽  
Author(s):  
M.M. Afifi ◽  
H.M. Atta ◽  
A.A. Elshanawan ◽  
U.M. Abdoul-rao ◽  
A.M. El-Adly
Keyword(s):  
Hygromycin B

Hygromycin B resistance as dominant selectable marker in yeast

1984 ◽  
Vol 8 (5) ◽  
pp. 353-358 ◽  
Author(s):  
Kevin R. Kaster ◽  
Stanley G. Burgett ◽  
Thomas D. Ingolia
Keyword(s):  
Selectable Marker ◽  
Hygromycin B ◽  
Hygromycin B Resistance ◽  
Dominant Selectable Marker

Dominant positive and negative selection using a hygromycin phosphotransferase-thymidine kinase fusion gene

1991 ◽  
Vol 11 (6) ◽  
pp. 3374-3378 ◽  
Author(s):  
S D Lupton ◽  
L L Brunton ◽  
V A Kalberg ◽  
R W Overell
Keyword(s):  
Thymidine Kinase ◽  
Negative Selection ◽  
Fusion Gene ◽  
Hygromycin B ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Hygromycin Phosphotransferase ◽  
Simplex Virus ◽  
Genetic Entity

The hygromycin phosphotransferase gene was fused in-frame with the herpes simplex virus type 1 thymidine kinase gene. The resulting fusion gene (termed HyTK) confers hygromycin B resistance for dominant positive selection and ganciclovir sensitivity for negative selection and provides a means by which these selectable phenotypes may be expressed and regulated as a single genetic entity.

Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells

1984 ◽  
Vol 4 (12) ◽  
pp. 2929-2931 ◽  
Author(s):  
K Blochlinger ◽  
H Diggelmann
Keyword(s):  
Selectable Marker ◽  
Dna Transfer ◽  
Direct Selection ◽  
Regulatory Sequences ◽  
Hygromycin B ◽  
Dominant Marker ◽  
Dna Coding ◽  
Sarcoma Virus ◽  
Moloney Sarcoma Virus ◽  
Selection For

The DNA coding sequence for the hygromycin B phosphotransferase gene was placed under the control of the regulatory sequences of a cloned long terminal repeat of Moloney sarcoma virus. This construction allowed direct selection for hygromycin B resistance after transfection of eucaryotic cell lines not naturally resistant to this antibiotic, thus providing another dominant marker for DNA transfer in eucaryotic cells.

Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay

Gene ◽  
1992 ◽  
Vol 112 (2) ◽  
pp. 257-260 ◽  
Author(s):  
Schandorf Sørensen Michael ◽  
Mogens Duch ◽  
Kirsten Paludan ◽  
Poul Jøgensen ◽  
Skou Pedersen Finn
Keyword(s):  
Mammalian Cell ◽  
Hygromycin B ◽  
Dot Blot ◽  
Cell Extracts ◽  
Dot Blot Assay

scholarly journals Studying the Optimum Conditions of ‎Hygromycin B Production and Detect their ‎Toxicity

2017 ◽  
Vol 26 (2) ◽  
pp. 119-130
Author(s):  
Zahraa Abdulmunim ‎ ◽  
Rabah N. Jabba ◽  
Abdulwahid B. Al-Shaibani
Keyword(s):  
Antimicrobial Activity ◽  
Aqueous Phase ◽  
Crude Extract ◽  
Incubation Temperature ◽  
Salmonella Typhi ◽  
Broth Culture ◽  
Hygromycin B ◽  
Yeast Saccharomyces Cerevisiae ◽  
Diffusion Technique ◽  
Mice Liver

Hygromycin B was extracted with ethyl acetate, which separates organic phase from aqueous phase in the broth culture filtrate, only the aqueous phase showed significant antimicrobial activity by using agar well diffusion technique. At a concentration of 25mg/ml (as crude extract), this phase excreted its activity against the test microorganisms which include; one G(+) bacteria (Staphylococcus aureus), five G(–) bacteria (Pseudomonas aeruginosa , Proteus mirabilis, Escherichia coli , Klebsiella pneumoniae, Salmonella typhi) and one yeast (Saccharomyces  cerevisiae).          After detecting the aminoglycoside hygromycin B by the Thin Layer Chromatography (TLC) method to ensure presence of the antibiotic, same flow rate (Rf) value (0.357) as that of the standard hygromycin B was obtained.          Results of the optimization conditions showed that the highest antimicrobial activity of hygromycin B was obtained at a medium pH of 8 and incubation temperature of 35°C for 10 days. When the toxicity of hygromycin B crude extract under such conditions was examined on mice liver, a mild effects were appeared

scholarly journals d-Sedoheptulose-7-phosphate is a common precursor for the heptoses of septacidin and hygromycin B

2018 ◽  
Vol 115 (11) ◽  
pp. 2818-2823 ◽  
Author(s):  
Wei Tang ◽  
Zhengyan Guo ◽  
Zhenju Cao ◽  
Min Wang ◽  
Pengwei Li ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Carbon Chain ◽  
Side Chain ◽  
Hygromycin B ◽  
Nucleoside Antibiotics ◽  
Common Precursor ◽  
Encoding Genes ◽  
Poultry Farming

Seven-carbon-chain–containing sugars exist in several groups of important bacterial natural products. Septacidin represents a group of l-heptopyranoses containing nucleoside antibiotics with antitumor, antifungal, and pain-relief activities. Hygromycin B, an aminoglycoside anthelmintic agent used in swine and poultry farming, represents a group of d-heptopyranoses–containing antibiotics. To date, very little is known about the biosynthesis of these compounds. Here we sequenced the genome of the septacidin producer and identified the septacidin gene cluster by heterologous expression. After determining the boundaries of the septacidin gene cluster, we studied septacidin biosynthesis by in vivo and in vitro experiments and discovered that SepB, SepL, and SepC can convert d-sedoheptulose-7-phosphate (S-7-P) to ADP-l-glycero-β-d-manno-heptose, exemplifying the involvement of ADP-sugar in microbial natural product biosynthesis. Interestingly, septacidin, a secondary metabolite from a gram-positive bacterium, shares the same ADP-heptose biosynthesis pathway with the gram-negative bacterium LPS. In addition, two acyltransferase-encoding genes sepD and sepH, were proposed to be involved in septacidin side-chain formation according to the intermediates accumulated in their mutants. In hygromycin B biosynthesis, an isomerase HygP can recognize S-7-P and convert it to ADP-d-glycero-β-d-altro-heptose together with GmhA and HldE, two enzymes from the Escherichia coli LPS heptose biosynthetic pathway, suggesting that the d-heptopyranose moiety of hygromycin B is also derived from S-7-P. Unlike the other S-7-P isomerases, HygP catalyzes consecutive isomerizations and controls the stereochemistry of both C2 and C3 positions.

Development of a fungal transformation system based on selection of sequences with promoter activity

1987 ◽  
Vol 7 (9) ◽  
pp. 3297-3305
Author(s):  
B G Turgeon ◽  
R C Garber ◽  
O C Yoder
Keyword(s):  
Genomic Dna ◽  
Pathogenic Fungus ◽  
Low Frequency ◽  
Hygromycin B ◽  
Fungal Transformation ◽  
Dna Fragments ◽  
Transformation System ◽  
Chromosomal Dna ◽  
General Utility ◽  
Novel Strategy

A novel strategy was used to develop a transformation system for the plant pathogenic fungus Cochliobolus heterostrophus. Sequences capable of driving the expression of a gene conferring resistance to the antibiotic hygromycin B in C. heterostrophus were selected from a library of genomic DNA fragments and used, with the selectable marker, as the basis for transformation. The library of random 0.5- to 2.0-kilobase-pair fragments of C. heterostrophus genomic DNA was inserted at the 5' end of a truncated, promoterless Escherichia coli hygromycin B phosphotransferase gene (hygB) whose product confers resistance to hygromycin B. C. heterostrophus protoplasts were transformed with the library and selected for resistance. Resistant colonies arose at low frequency. Each colony contained a transformation vector stably integrated into chromosomal DNA. When the transforming DNA was recovered from the genome and introduced into C. heterostrophus, resistant colonies appeared at higher frequency. We determined the sequences of two of the C. heterostrophus DNA fragments which had been inserted at the 5' end of hygB in the promoter library and found that both made translational fusions with hygB. One of the two fusions apparently adds 65 and the other at least 86 amino acids to the N-terminus of the hygB product. Plasmids containing hygB-C. heterostrophus promoter fusions can be used unaltered to drive hygB expression in several other filamentous ascomycetes. This approach to achieving transformation may have general utility, especially for organisms with relatively undeveloped genetics.

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