scholarly journals Isolation and Characterization of Insecticidal Toxins from the Venom of the North African Scorpion, Buthacus leptochelys

Toxins ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 236 ◽  
Author(s):  
Yusuke Yoshimoto ◽  
Masahiro Miyashita ◽  
Mohammed Abdel-Wahab ◽  
Moustafa Sarhan ◽  
Yoshiaki Nakagawa ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Scorpion Venom ◽  
Spectrometric Analysis ◽  
North African ◽  
High Sequence Similarity ◽  
The North ◽  
Isolation And Characterization ◽  
Ms Analysis ◽  
Insect Toxicity

Various bioactive peptides have been identified in scorpion venom, but there are many scorpion species whose venom has not been investigated. In this study, we characterized venom components of the North African scorpion, Buthacus leptochelys, by mass spectrometric analysis and evaluated their insect toxicity. This is the first report of chemical and biological characterization of the B. leptochelys venom. LC/MS analysis detected at least 148 components in the venom. We isolated four peptides that show insect toxicity (Bl-1, Bl-2, Bl-3, and Bl-4) through bioassay-guided HPLC fractionation. These toxins were found to be similar to scorpion α- and β-toxins based on their N-terminal sequences. Among them, the complete primary structure of Bl-1 was determined by combination of Edman degradation and MS/MS analysis. Bl-1 is composed of 67 amino acid residues and crosslinked with four disulfide bonds. Since Bl-1 shares high sequence similarity with α-like toxins, it is likely that it acts on Na+ channels of both insects and mammals.

scholarly journals The embryonic RNA helicase gene (ERH): a new member of the DEAD box family of RNA helicases

1995 ◽  
Vol 308 (3) ◽  
pp. 839-846 ◽  
Author(s):  
J Sowden ◽  
W Putt ◽  
K Morrison ◽  
R Beddington ◽  
Y Edwards
Keyword(s):  
Expression Profile ◽  
Sequence Similarity ◽  
Rna Helicase ◽  
Chromosome 1 ◽  
Rna Helicases ◽  
High Sequence Similarity ◽  
Dead Box ◽  
Isolation And Characterization ◽  
Helicase Gene ◽  
The Dead

DEAD box proteins share several highly conserved motifs including the characteristic Asp-Glu-Ala-Asp (D-E-A-D in the amino acid single-letter code) motif and have established or putative ATP-dependent RNA helicase activity. These proteins are implicated in a range of cellular processes that involve regulation of RNA function, including translation initiation, RNA splicing and ribosome assembly. Here we describe the isolation and characterization of an embryonic RNA helicase gene, ERH, which maps to mouse chromosome 1 and encodes a new member of the DEAD box family of proteins. The predicted ERH protein shows high sequence similarity to the testes-specific mouse PL10 and to the maternally acting Xenopus An3 helicase proteins. The ERH expression profile is similar, to that of An3, which localizes to the animal hemisphere of oocytes and is abundantly expressed in the embryo. ERH is expressed in oocytes and is a ubiquitous mRNA in the 9 days-post-conception embryo, and at later stages of development shows a more restricted pattern of expression in brain and kidney. The similarities in sequence and in expression profile suggest that ERH is the murine equivalent of the Xenopus An3 gene, and we propose that ERH plays a role in translational activation of mRNA in the oocyte and early embryo.

scholarly journals Isolation and Characterization of the Gene Coding for the Amidinotransferase Involved in the Biosynthesis of Phaseolotoxin in Pseudomonas syringae pv. phaseolicola

2001 ◽  
Vol 14 (4) ◽  
pp. 545-554 ◽  
Author(s):  
Gustavo Hernández-Guzmán ◽  
Ariel Alvarez-Morales
Keyword(s):  
Aspartic Acid ◽  
Pseudomonas Syringae ◽  
Sequence Similarity ◽  
Insertion Mutant ◽  
Halo Blight ◽  
Streptomyces Spp ◽  
Isolation And Characterization ◽  
Gene Coding ◽  
Blight Disease

Pseudomonas syringae pv. phaseolicola is the causal agent of the “halo blight” disease of beans. A key component in the development of the disease is a nonhost-specific toxin, Nδ-(N'-sulphodiaminophosphinyl)-ornithyl-alanyl-homoarginine, known as phaseolotoxin. The homoarginine residue in this molecule has been suggested to be the product of Larginine:lysine amidinotransferase activity, previously detected in extracts of P. syringae pv. phaseolicola grown under conditions of phaseolotoxin production. We report the isolation and characterization of an amidinotransferase gene (amtA) from P. syringae pv. phaseolicola coding for a polypeptide of 362 residues (41.36 kDa) and showing approximately 40% sequence similarity to Larginine:inosamine-phosphate amidinotransferase from three species of Streptomyces spp. and 50.4% with an Larginine:glycine amidinotransferase from human mitochondria. The cysteine, histidine, and aspartic acid residues involved in substrate binding are conserved. Furthermore, expression of the amtA and argK genes and phaseolotoxin production occurs at 18°C but not at 28°C. An amidinotransferase insertion mutant was obtained that lost the capacity to synthesize homoarginine and phaseolotoxin. These results show that the amtA gene isolated is responsible for the amidinotransferase activity detected previously and that phaseolotoxin production depends upon the activity of this gene.

Isolation and Characterization of Soil Myxobacteria from Nepal

2019 ◽  
Vol 24 (2) ◽  
pp. 7-16
Author(s):  
Nabin Rana ◽  
Saraswoti Khadka ◽  
Bishnu Prasad Marasini ◽  
Bishnu Joshi ◽  
Pramod Poudel ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Antimicrobial Properties ◽  
Antimicrobial Activities ◽  
Rrna Gene ◽  
Isolation And Characterization ◽  
The Family ◽  
Global Concern ◽  
Bacteria And Fungi ◽  
Antimicrobial Metabolites

 Realizing myxobacteria as a potential source of antimicrobial metabolites, we pursued research to isolate myxobacteria showing antimicrobial properties. We have successfully isolated three strains (NR-1, NR-2, NR-3) using the Escherichia coli baiting technique. These isolates showed typical myxobacterial growth characteristics. Phylogenetic analysis showed that all the strains (NR-1, NR-2, NR-3) belong to the family Archangiaceae, suborder Cystobacterineae, and order Myxococcales. Furthermore, 16S rRNA gene sequence similarity searched through BLAST revealed that strain NR-1 showed the closest similarity (91.8 %) to the type strain Vitiosangium cumulatum (NR-156939), NR-2 showed (98.8 %) to the type of Cystobacter badius (NR-043940), and NR-3 showed the closest similarity (83.5 %) to the type of strain Cystobacter fuscus (KP-306730). All isolates showed better growth in 0.5-1 % NaCl and pH around 7.0, whereas no growth was observed at pH 9.0 and below 5.0. All strains showed better growth at 32° C and hydrolyzed starch, whereas casein was efficiently hydrolyzed by NR-1 and NR-2. Besides, preliminary antimicrobial tests from crude extracts showed activities against Gram-positive, Gram-negative bacteria, and fungi. Our findings suggest that the arcane soil habitats of Nepal harbor myxobacteria with the capability to produce diverse antimicrobial activities that may be explored to overcome the rapidly rising global concern about antibiotic resistance.

scholarly journals Isolation and Characterization of Novel Psychrophilic, Neutrophilic, Fe-Oxidizing, Chemolithoautotrophic α- and γ-Proteobacteria from the Deep Sea

2003 ◽  
Vol 69 (5) ◽  
pp. 2906-2913 ◽  
Author(s):  
K. J. Edwards ◽  
D. R. Rogers ◽  
C. O. Wirsen ◽  
T. M. McCollom
Keyword(s):  
Deep Sea ◽  
Sequence Similarity ◽  
Heterotrophic Bacterium ◽  
Physiological Characterization ◽  
Metalliferous Sediments ◽  
Basalt Glass ◽  
Isolation And Characterization ◽  
Iron Oxidizing Bacteria ◽  
Microaerobic Conditions

ABSTRACT We report the isolation and physiological characterization of novel, psychrophilic, iron-oxidizing bacteria (FeOB) from low-temperature weathering habitats in the vicinity of the Juan de Fuca deep-sea hydrothermal area. The FeOB were cultured from the surfaces of weathered rock and metalliferous sediments. They are capable of growth on a variety of natural and synthetic solid rock and mineral substrates, such as pyrite (FeS2), basalt glass (∼10 wt% FeO), and siderite (FeCO3), as their sole energy source, as well as numerous aqueous Fe substrates. Growth temperature characteristics correspond to the in situ environmental conditions of sample origin; the FeOB grow optimally at 3 to 10°C and at generation times ranging from 57 to 74 h. They are obligate chemolithoautotrophs and grow optimally under microaerobic conditions in the presence of an oxygen gradient or anaerobically in the presence of nitrate. None of the strains are capable of using any organic or alternate inorganic substrates tested. The bacteria are phylogenetically diverse and have no close Fe-oxidizing or autotrophic relatives represented in pure culture. One group of isolates are γ-Proteobacteria most closely related to the heterotrophic bacterium Marinobacter aquaeolei (87 to 94% sequence similarity). A second group of isolates are α-Proteobacteria most closely related to the deep-sea heterotrophic bacterium Hyphomonas jannaschiana (81 to 89% sequence similarity). This study provides further evidence for the evolutionarily widespread capacity for Fe oxidation among bacteria and suggests that FeOB may play an unrecognized geomicrobiological role in rock weathering in the deep sea.

scholarly journals Tuning the ion selectivity of two-pore channels

2017 ◽  
Vol 114 (5) ◽  
pp. 1009-1014 ◽  
Author(s):  
Jiangtao Guo ◽  
Weizhong Zeng ◽  
Youxing Jiang
Keyword(s):  
Sequence Similarity ◽  
Ion Selectivity ◽  
Structural Basis ◽  
Monovalent Cations ◽  
Nonselective Cation Channel ◽  
High Sequence Similarity ◽  
Selective Channel ◽  
Tm Domain ◽  
Key Residues

Organellar two-pore channels (TPCs) contain two copies of aShaker-like six-transmembrane (6-TM) domain in each subunit and are ubiquitously expressed in plants and animals. Interestingly, plant and animal TPCs share high sequence similarity in the filter region, yet exhibit drastically different ion selectivity. Plant TPC1 functions as a nonselective cation channel on the vacuole membrane, whereas mammalian TPC channels have been shown to be endo/lysosomal Na+-selective or Ca2+-release channels. In this study, we performed systematic characterization of the ion selectivity of TPC1 fromArabidopsis thaliana(AtTPC1) and compared its selectivity with the selectivity of human TPC2 (HsTPC2). We demonstrate that AtTPC1 is selective for Ca2+over Na+, but nonselective among monovalent cations (Li+, Na+, and K+). Our results also confirm that HsTPC2 is a Na+-selective channel activated by phosphatidylinositol 3,5-bisphosphate. Guided by our recent structure of AtTPC1, we converted AtTPC1 to a Na+-selective channel by mimicking the selectivity filter of HsTPC2 and identified key residues in the TPC filters that differentiate the selectivity between AtTPC1 and HsTPC2. Furthermore, the structure of the Na+-selective AtTPC1 mutant elucidates the structural basis for Na+selectivity in mammalian TPCs.

scholarly journals Isolation and Characterization of Endophytic Bacteria from Tembelekan (Lantana camara L.) as Antibacterial Compounds Producer

2015 ◽  
Vol 2 (2) ◽  
pp. 86-98
Author(s):  
Dina Dyah Saputri ◽  
Maria Bintang ◽  
Fachriyan H Pasaribu
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Endophytic Bacteria ◽  
Pathogenic Bacteria ◽  
Lantana Camara ◽  
Rrna Gene ◽  
Antibacterial Compounds ◽  
Isolation And Characterization ◽  
Ms Analysis

Endophytic bacteria are microorganisms that live in the internal tissues of plants and have symbiotic mutualism with their host plants. Endophytic bacteria may produce secondary metabolites that can be developed for medical, agricultural, and industrial purposes. Lantana camara is a medicinal plant that has therapeutic potential to treat a variety of diseases such as fever, tuberculosis, rheumatism, asthma, and skin disease. The purpose of this study was to isolate and characterize endophytic bacteria from Lantana camara which has potential to produce antibacterial compounds. The method of this research include isolation of endophytic bacteria of Lantana camara. Antibacterial activity assay was done against four types of pathogenic bacteria i.e. Bacillus cereus, Escherichia coli, Staphylococcus aureus, and Salmonella enteritidis. Characterization of endophytic bacteria was by 16S rRNA gene analysis and identification of antibacterial compounds by GC-MS analysis. Isolation of endophytic bacteria from Lantana camara resulted in BT22 as a potential isolate. Analysis of 16S rRNA gene showed that the BT22 isolate was similar to Bacillus amyloliquefaciens YB-1402 with 99% identity. The results of GC-MS analysis showed some antibacterial compounds such as: Cyclohexanone, 2-[2-(1,3-dithiolan-2-yl)propyl]-6-methyl-3-(1-methylethyl), Octadecane (CAS) n-Octadecane and Tetracosane (CAS) n-Tetracosane.

scholarly journals Inheritance of seedlessness and the molecular characterization of the INO gene in Annonaceae

2023 ◽  
Vol 83 ◽  
Author(s):  
B. R. R. M. Nassau ◽  
P. S. C. Mascarenhas ◽  
A. G. Guimarães ◽  
F. M. Feitosa ◽  
H. M. Ferreira ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Type A ◽  
Molecular Techniques ◽  
Wild Type ◽  
Annona Squamosa ◽  
High Sequence Similarity ◽  
Breeding Programs ◽  
Complete Dominance ◽  
Young Leaves

Abstract The inheritance of the seedless fruit characteristic of Annona squamosa has not yet been explained. Molecular techniques may aid breeding programs, mainly in the assisted selection of the target gene. The INO gene may be related to seed development in these fruits. The objective of the present paper was to investigate the inheritance of seedlessness in the 'Brazilian seedless' sugar apple and INO gene conservation in Annona squamosa and Annona cherimola x Annona squamosa genotypes by assessing their homology with the INO database genes. The F1 generation was obtained by crossing the mutant 'Brazilian seedless' (male genitor) (P1) with the wild-type A. squamosa with seeds (M1 and M2, female genitors). The INO gene was studied in mutant and wild-type A. squamosa (P1, M1, M2 and M3) and in the Gefner atemoya (A. cherimola x A. squamosa) (M4) cultivar. The DNA was extracted from young leaves, and four sets of specific primers flanking the INO gene were amplified. The seedless characteristic was identified as stenospermatic in the fruits of parental P1, suggesting monogenic inheritance with complete dominance. High sequence similarity of the INO gene amplifications in the sugar apple accessions (M1, M2, M3) and the atemoya cultivar Gefner (M4) reinforces the hypothesis of their conservation.

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