scholarly journals Identification of Methyl Halide-Utilizing Genes in the Methyl Bromide-Utilizing Bacterial Strain IMB-1 Suggests a High Degree of Conservation of Methyl Halide-Specific Genes in Gram-Negative Bacteria

2001 ◽  
Vol 67 (4) ◽  
pp. 1959-1963 ◽  
Author(s):  
Claire A. Woodall ◽  
Karen L. Warner ◽  
Ronald S. Oremland ◽  
J. Colin Murrell ◽  
Ian R. McDonald
Keyword(s):  
Energy Source ◽  
Methyl Bromide ◽  
Open Reading Frames ◽  
Binding Motif ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
High Sequence Homology ◽  
Methyl Halide ◽  
High Degree ◽  
Reading Frames

ABSTRACT Strain IMB-1, an aerobic methylotrophic member of the alpha subgroup of the Proteobacteria, can grow with methyl bromide as a sole carbon and energy source. A single cmugene cluster was identified in IMB-1 that contained six open reading frames: cmuC, cmuA, orf146, paaE, hutI, and partialmetF. CmuA from IMB-1 has high sequence homology to the methyltransferase CmuA from Methylobacterium chloromethanicum and Hyphomicrobium chloromethanicum and contains a C-terminal corrinoid-binding motif and an N-terminal methyltransferase motif. However,cmuB, identified in M. chloromethanicumand H. chloromethanicum, was not detected in IMB-1.

scholarly journals All Seven comG Open Reading Frames Are Required for DNA Binding during Transformation of CompetentBacillus subtilis

1998 ◽  
Vol 180 (1) ◽  
pp. 41-45 ◽  
Author(s):  
Y. S. Chung ◽  
D. Dubnau
Keyword(s):  
Bacillus Subtilis ◽  
Dna Binding ◽  
Cell Surface ◽  
Open Reading Frames ◽  
Gram Negative Bacteria ◽  
Gene Products ◽  
Gram Negative ◽  
Transposon Insertions ◽  
Reading Frames

ABSTRACT The seven proteins encoded by the comG operon ofBacillus subtilis exhibit similarity to gene products required for the assembly of type 4 pili and for the secretion of certain proteins in gram-negative bacteria. Although polar transposon insertions in comG result in the loss of transformability and in the failure of cells grown through the competence regimen to bind DNA, it was not known whether the ComG proteins are all required for competence. We have constructed strains missing each of these proteins individually and found that they are all nontransformable and fail to bind transforming DNA to the cell surface. The implications of these findings are discussed.

scholarly journals Two-Partner Secretion Systems of Neisseria meningitidis Associated with Invasive Clonal Complexes

2008 ◽  
Vol 76 (10) ◽  
pp. 4649-4658 ◽  
Author(s):  
Peter van Ulsen ◽  
Lucy Rutten ◽  
Moniek Feller ◽  
Jan Tommassen ◽  
Arie van der Ende
Keyword(s):  
Neisseria Meningitidis ◽  
Outer Membrane ◽  
Open Reading Frames ◽  
Gram Negative Bacteria ◽  
Secretion Systems ◽  
Gram Negative ◽  
System 1 ◽  
Related Proteins ◽  
Encoding Genes ◽  
Reading Frames

ABSTRACT The two-partner secretion (TPS) pathway is widespread among gram-negative bacteria and facilitates the secretion of very large and often virulence-related proteins. TPS systems consist of a secreted TpsA protein and a TpsB protein involved in TpsA transport across the outer membrane. Sequenced Neisseria meningitidis genomes contain up to five TpsA- and two TpsB-encoding genes. Here, we investigated the distribution of TPS-related open reading frames in a collection of disease isolates. Three distinct TPS systems were identified among meningococci. System 1 was ubiquitous, while systems 2 and 3 were significantly more prevalent among isolates of hyperinvasive clonal complexes than among isolates of poorly invasive clonal complexes. In laboratory cultures, systems 1 and 2 were expressed. However, several sera from patients recovering from disseminated meningococcal disease recognized the TpsAs of systems 2 and 3, indicating the expression of these systems during infection. Furthermore, we showed that the major secreted TpsAs of systems 1 and 2 depend on their cognate TpsBs for transport across the outer membrane and that the system 1 TpsAs undergo processing. Together, our data indicate that TPS systems may contribute to the virulence of N. meningitidis.

scholarly journals Sequence of the Genome of the Temperate, Serotype-Converting,Pseudomonas aeruginosa Bacteriophage D3

2000 ◽  
Vol 182 (21) ◽  
pp. 6066-6074 ◽  
Author(s):  
Andrew M. Kropinski
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Sequence Similarity ◽  
Complete Sequence ◽  
Open Reading Frames ◽  
Gene Encoding ◽  
Virus Family ◽  
Double Stranded Dna ◽  
High Degree ◽  
Phage Proteins ◽  
Reading Frames

ABSTRACT Temperate bacteriophage D3, a member of the virus familySiphoviridae, is responsible for serotype conversion in its host, Pseudomonas aeruginosa. The complete sequence of the double-stranded DNA genome has been determined. The 56,426 bp contains 90 putative open reading frames (ORFs) and four genes specifying tRNAs. The latter are specific for methionine (AUG), glycine (GGA), asparagine (AAC), and threonine (ACA). The tRNAs may function in the translation of certain highly expressed proteins from this relatively AT-rich genome. D3 proteins which exhibited a high degree of sequence similarity to previously characterized phage proteins included the portal, major head, tail, and tail tape measure proteins, endolysin, integrase, helicase, and NinG. The layout of genes was reminiscent of lambdoid phages, with the exception of the placement of the endolysin gene, which parenthetically also lacked a cognate holin. The greatest sequence similarity was found in the morphogenesis genes to coliphages HK022 and HK97. Among the ORFs was discovered the gene encoding the fucosamine O-acetylase, which is in part responsible for the serotype conversion events.

scholarly journals Sequence and Organization of pXO1, the Large Bacillus anthracis Plasmid Harboring the Anthrax Toxin Genes

1999 ◽  
Vol 181 (20) ◽  
pp. 6509-6515 ◽  
Author(s):  
R. T. Okinaka ◽  
K. Cloud ◽  
O. Hampton ◽  
A. R. Hoffmaster ◽  
K. K. Hill ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Regulatory Elements ◽  
Open Reading Frames ◽  
Significant Similarity ◽  
Toxin Genes ◽  
Theta Replication ◽  
Large Plasmids ◽  
High Degree ◽  
Degree Of Similarity ◽  
Reading Frames

ABSTRACT The Bacillus anthracis Sterne plasmid pXO1 was sequenced by random, “shotgun” cloning. A circular sequence of 181,654 bp was generated. One hundred forty-three open reading frames (ORFs) were predicted using GeneMark and GeneMark.hmm, comprising only 61% (110,817 bp) of the pXO1 DNA sequence. The overall guanine-plus-cytosine content of the plasmid is 32.5%. The most recognizable feature of the plasmid is a “pathogenicity island,” defined by a 44.8-kb region that is bordered by inverted IS1627 elements at each end. This region contains the three toxin genes (cya, lef, and pagA), regulatory elements controlling the toxin genes, three germination response genes, and 19 additional ORFs. Nearly 70% of the ORFs on pXO1 do not have significant similarity to sequences available in open databases. Absent from the pXO1 sequence are homologs to genes that are typically required to drive theta replication and to maintain stability of large plasmids in Bacillus spp. Among the ORFs with a high degree of similarity to known sequences are a collection of putative transposases, resolvases, and integrases, suggesting an evolution involving lateral movement of DNA among species. Among the remaining ORFs, there are three sequences that may encode enzymes responsible for the synthesis of a polysaccharide capsule usually associated with serotype-specific virulent streptococci.

scholarly journals Physicochemical and Antibacterial Properties of Chitosan Extracted from Waste Shrimp Shells

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
José Carlos Vilar Junior ◽  
Daylin Rubio Ribeaux ◽  
Carlos Alberto Alves da Silva ◽  
Galba Maria De Campos-Takaki
Keyword(s):  
Antibacterial Activity ◽  
Antibacterial Properties ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Degree Of Deacetylation ◽  
Shrimp Shells ◽  
Shrimp Shell ◽  
Ft Ir ◽  
High Degree

This research aims to study the production of chitosan from shrimp shell (Litopenaeus vannamei) of waste origin using two chemical methodologies involving demineralization, deproteinization, and the degree of deacetylation. The evaluation of the quality of chitosan from waste shrimp shells includes parameters for the yield, physical chemistry characteristics by infrared spectroscopy (FT-IR), the degree of deacetylation, and antibacterial activity. The results showed (by Method 1) extraction yields for chitin of 33% and for chitosan of 49% and a 76% degree of deacetylation. Chitosan obtained by Method 2 was more efficient: chitin (36%) and chitosan (63%), with a high degree of deacetylation (81.7%). The antibacterial activity was tested against Gram-negative bacteria (Stenotrophomonas maltophiliaandEnterobacter cloacae) and Gram-positiveBacillus subtilisand the Minimum Inhibitory Concentrations (MIC) and the Minimum Bactericidal Concentration (MBC) were determined. Method 2 showed that extracted chitosan has good antimicrobial potential against Gram-positive and Gram-negative bacteria and that the process is viable.

scholarly journals An O-Phosphotransferase Catalyzes Phosphorylation of Hygromycin A in the Antibiotic-Producing Organism Streptomyces hygroscopicus

2008 ◽  
Vol 52 (10) ◽  
pp. 3580-3588 ◽  
Author(s):  
Vidya Dhote ◽  
Shuchi Gupta ◽  
Kevin A. Reynolds
Keyword(s):  
Ribosomal Subunit ◽  
Biosynthetic Gene Cluster ◽  
Open Reading Frames ◽  
Streptomyces Hygroscopicus ◽  
Gram Negative Bacteria ◽  
E Coli ◽  
Naturally Occurring ◽  
Insertional Inactivation ◽  
Pathway Regulation ◽  
Reading Frames

ABSTRACT The antibiotic hygromycin A (HA) binds to the 50S ribosomal subunit and inhibits protein synthesis in gram-positive and gram-negative bacteria. The HA biosynthetic gene cluster in Streptomyces hygroscopicus NRRL 2388 contains 29 open reading frames, which have been assigned putative roles in biosynthesis, pathway regulation, and self-resistance. The hyg21 gene encodes an O-phosphotransferase with a proposed role in self-resistance. We observed that insertional inactivation of hyg21 in S. hygroscopicus leads to a greater than 90% decrease in HA production. The wild type and the hyg21 mutant were comparably resistant to HA. Using Escherichia coli as a heterologous host, we expressed and purified Hyg21. Kinetic analyses revealed that the recombinant protein catalyzes phosphorylation of HA (Km = 30 ± 4 μM) at the C-2‴ position of the fucofuranose ring in the presence of ATP (Km = 200 ± 20 μM) or GTP (Km = 350 ± 60 μM) with a k cat of 2.2 ± 0.1 min−1. The phosphorylated HA is inactive against HA-sensitive ΔtolC E. coli and Streptomyces lividans. Hyg21 also phosphorylates methoxyhygromycin A and desmethylenehygromycin A with k cat and Km values similar to those observed with HA. Phosphorylation of the naturally occurring isomers of 5‴-dihydrohygromycin A and 5‴-dihydromethoxyhygromycin A was about 12 times slower than for the corresponding non-natural isomers. These studies demonstrate that Hyg21 is an O-phosphotransferase with broad substrate specificity, tolerating changes in the aminocyclitol moiety more than in the fucofuranose moiety, and that phosphorylation by Hyg21 is one of several possible mechanisms of self-resistance in S. hygroscopicus NRRL 2388.

A WIDE-SPECTRUM ANTIBIOTIC PRODUCED BY A SPECIES OF SORANGIUM

1966 ◽  
Vol 12 (2) ◽  
pp. 221-230 ◽  
Author(s):  
E. A. Peterson ◽  
D. C. Gillespie ◽  
F. D. Cook
Keyword(s):  
Low Temperature ◽  
Aqueous Solutions ◽  
Phosphate Buffer ◽  
Wide Spectrum ◽  
Active Material ◽  
Gram Negative Bacteria ◽  
Alkaline Ph ◽  
Gram Negative ◽  
Ether Extraction ◽  
High Degree

A soil-borne myxobacter identified as a species of Sorangium produced a potent antibiotic capable of inhibiting growth of a wide variety of microorganisms including Gram-positive and Gram-negative bacteria, fungi, actinomycetes, and yeasts. The active material was readily isolated from culture fluids of the organism by ether extraction or by adsorption on a resin. A high degree of purity was achieved chromatographically. Acetone, methanol, or aqueous solutions of the antibiotic were stable when stored at low temperature (4 °C). At 70 °C it was unstable in phosphate buffer but retained its activity in iris buffer at neutral and alkaline pH.

scholarly journals Transgenic Kalanchoë blossfeldiana, Containing Individual rol Genes and Open Reading Frames Under 35S Promoter, Exhibit Compact Habit, Reduced Plant Growth, and Altered Ethylene Tolerance in Flowers

2021 ◽  
Vol 12 ◽  
Author(s):  
Bruno Trevenzoli Favero ◽  
Yi Tan ◽  
Yan Lin ◽  
Hanne Bøge Hansen ◽  
Nasim Shadmani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Plant Growth ◽  
Growth Habit ◽  
Open Reading Frames ◽  
Gene Copy ◽  
Binding Motif ◽  
Common Denominator ◽  
Kalanchoe Blossfeldiana ◽  
Reading Frames ◽  
Kalanchoë Blossfeldiana

Reduced growth habit is a desirable trait for ornamental potted plants and can successfully be obtained through Rhizobium rhizogenes transformation in a stable and heritable manner. Additionally, it can also be obtained by transformation with Agrobacterium tumefaciens harboring specific genes from R. rhizogenes. The bacterial T-DNA harbors four root oncogenic loci (rol) genes and 14 less known open reading frames (ORFs). The four rol genes, i.e., rolA, rolB, rolC, and rolD, are conceived as the common denominator for the compact phenotype and the other less characterized ORFs seem auxiliary but present a potential breeding target for less aberrant and/or more tailored phenotypes. In this study, Kalanchoë blossfeldiana ‘Molly’ was transformed with individual rol genes and selected ORFs in 35S overexpressing cassettes to comprehensively characterize growth traits, gene copy and expression, and ethylene tolerance of the flowers. An association of reduced growth habit, e.g. height and diameter, was observed for rolB2 and ORF14-2 when a transgene single copy and high gene expression were detected. Chlorophyll content was reduced in overexpressing lines compared to wild type (WT), except for one ΔORF13a (a truncated ORF13a, where SPXX DNA-binding motif is absent). The flower number severely decreased in the overexpressing lines compared to WT. The anthesis timing showed that WT opened the first flower at 68.9 ± 0.9 days and the overexpressing lines showed similar or up to 24 days delay in flowering. In general, a single or low relative gene copy insertion was correlated to higher gene expression, ca. 3 to 5-fold, in rolB and ΔORF13a lines, while in ORF14 such relation was not directly linked. The increased gene expression observed in rolB2 and ΔORF13a-2 contributed to reducing plant growth and a more compact habit. Tolerance of detached flowers to 0.5 μl L−1 ethylene was markedly higher for ORF14 with 66% less flower closure at day 3 compared to WT. The subcellular localization of rolC and ΔORF13a was investigated by transient expression in Nicotiana benthamiana and confocal images showed that rolC and ΔORF13a are soluble and localize in the cytoplasm being able to enter the nucleus.

scholarly journals Patterns of Sequence Conservation in the S-Layer Proteins and Related Sequences in Clostridium difficile

2002 ◽  
Vol 184 (14) ◽  
pp. 3886-3897 ◽  
Author(s):  
Emanuela Calabi ◽  
Neil Fairweather
Keyword(s):  
Clostridium Difficile ◽  
Open Reading Frames ◽  
C Terminus ◽  
Antibiotic Associated Diarrhea ◽  
Processing Site ◽  
Different Strains ◽  
Related Proteins ◽  
High Degree ◽  
Insight Into ◽  
Reading Frames

ABSTRACT Clostridium difficile is the etiological agent of antibiotic-associated diarrhea. Among the factors that may play a role in infection are S-layer proteins (SLPs). Previous work has shown these to consist mainly of two components, resulting from the cleavage of a precursor encoded by the slpA gene. The high-molecular-weight (MW) subunit is related both to amidases from B. subtilis and to at least another 28 gene products in C. difficile strain 630. To gain insight into the functions of the SLPs and related proteins, we have further investigated the pattern of variability both at the slpA locus and at six nearby paralogs. Sequencing of the slpA gene from an S-layer group II strain and a variant S-layer group strain confirms a high degree of divergence in the low-MW SLP, which may result from diversifying selection. A highly conserved motif, however, is found at the C terminus in all low-MW subunits and may be essential for SlpA precursor cleavage. In strain 167, a variant cleavage product is present, suggesting a secondary processing site. Southern blotting analysis shows slpA-like open reading frames (ORFs) 2 to 7 to be conserved in all nine strains tested, with one exception: ORF2, which encodes a 66-kDa polypeptide coextracted at low pH with the main SLPs in strain 630, may be partially deleted in strain 167. Polymorphism within the slpA-ORF7 cluster may be more pronounced in the region proximal to the slpA gene. Unexpectedly, a high-MW subunit probe cross hybridizes to sequences outside the slpA locus, which appear to vary in number in different strains.

scholarly journals How Bacteria Consume Their Own Exoskeletons (Turnover and Recycling of Cell Wall Peptidoglycan)

2008 ◽  
Vol 72 (2) ◽  
pp. 211-227 ◽  
Author(s):  
James T. Park ◽  
Tsuyoshi Uehara
Keyword(s):  
Energy Source ◽  
Degradation Products ◽  
Side Wall ◽  
Amino Sugars ◽  
Gram Negative Bacteria ◽  
Periplasmic Binding Protein ◽  
Gram Negative ◽  
E Coli ◽  
The Individual

SUMMARY The phenomenon of peptidoglycan recycling is reviewed. Gram-negative bacteria such as Escherichia coli break down and reuse over 60% of the peptidoglycan of their side wall each generation. Recycling of newly made peptidoglycan during septum synthesis occurs at an even faster rate. Nine enzymes, one permease, and one periplasmic binding protein in E. coli that appear to have as their sole function the recovery of degradation products from peptidoglycan, thereby making them available for the cell to resynthesize more peptidoglycan or to use as an energy source, have been identified. It is shown that all of the amino acids and amino sugars of peptidoglycan are recycled. The discovery and properties of the individual proteins and the pathways involved are presented. In addition, the possible role of various peptidoglycan degradation products in the induction of β-lactamase is discussed.

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