The puh structural gene coding for the H subunit of the Rhodospirillum rubrum photoreaction center

1991 ◽  
Vol 69 (2-3) ◽  
pp. 122-131 ◽  
Author(s):  
Jacques Bérard ◽  
Gabriel Gingras
Keyword(s):  
Structural Gene ◽  
Rhodospirillum Rubrum ◽  
Rhodobacter Capsulatus ◽  
Bacterial Species ◽  
Polyacrylamide Gel Electrophoresis ◽  
Open Reading Frames ◽  
The Other ◽  
Rhodopseudomonas Viridis ◽  
Glutamic Acid Residue ◽  
Contact Points

The Rhodospirillum rubrum structural gene puh, coding for the photoreaction center H polypeptide, and three other putative genes that surround puh were cloned and sequenced. The deduced 257 amino acid H polypeptide has a molecular weight of 27 909, in close agreement with polyacrylamide gel electrophoresis determination. Hydropathy plots predict a single hydrophobic α helix. The H polypeptide of Rhodospirillum rubrum shares only 23% of its residues with all three of the H polypeptides from Rhodopseudomonas viridis, Rhodobacter capsulatus, and Rhodobacter sphaeroides. Despite this apparent low degree of similarity, statistical analysis leaves no doubt about their close relatedness. Interspecies evolutionary distance, assessed by this analysis, confirms the closeness of the two Rhodobacter species, Rhodospirillum rubrum and Rhodopseudomonas viridis being approximately equidistant from them. Three regions of the H polypeptide are highly conserved in all four species. They correspond to known contact points of H with the complex of the other two (L and M) subunits on the cytoplasmic side of the membrane. A glutamic acid residue (H polypeptide residue 177), conserved in the other bacteria and suggested to be involved in the binding of secondary quinone QB, is replaced by serine in Rhodospirillum rubrum. The open reading frames G115, I2372, and I3087 are predicted to, respectively, encode polypeptides of 480, 224, and 155 residues coiled in 10, 2, and 1 transmembrane helices. Open reading frame G115 shares 56% identical residues with F1696, a sequence similarly arranged in the genome of Rhodobacter capsulatus. The gene product of ORF 13087 is predicted to share highly similar sequences with nitrogenase reductase (encoded by nifH) of 11 different bacterial species and is suggested to have a regulatory function.Key words: gene puh, DNA sequence, codon usage, evolution.

scholarly journals Molecular Cloning and Expression Analysis of the Rhodobacter capsulatus sodB Gene, Encoding an Iron Superoxide Dismutase

1998 ◽  
Vol 180 (20) ◽  
pp. 5413-5420 ◽  
Author(s):  
Néstor Cortez ◽  
Néstor Carrillo ◽  
Cécile Pasternak ◽  
Angelika Balzer ◽  
Gabriele Klug
Keyword(s):  
Superoxide Dismutase ◽  
Rhodobacter Capsulatus ◽  
Sequence Similarity ◽  
Bacterial Species ◽  
Polyacrylamide Gel Electrophoresis ◽  
Selection Procedure ◽  
Cloning And Expression ◽  
Gene Encoding ◽  
Reading Frame ◽  
Species Activity

ABSTRACT Genetic complementation of a sodA sodB Escherichia colimutant strain was used to clone Rhodobacter capsulatusgenes involved in detoxification of superoxide radicals. After sequence analysis, 1 of the 16 identical clones obtained by this selection procedure was shown to contain an open reading frame with sequence similarity to that coding for Fe-containing superoxide dismutases (SodB). The R. capsulatus sodB gene was expressed inE. coli, and the nature of the metal ligand was confirmed by inhibitor sensitivity assays with lysates from both bacterial species. Activity staining of cleared Rhodobacter lysates resolved by polyacrylamide gel electrophoresis indicated that SodB was the only superoxide dismutase present in this phototrophic organism. The sodB gene was expressed at low levels in R. capsulatus cells grown under anaerobic or semiaerobic conditions, but expression was strongly induced upon exposure of the bacteria to air or to methyl viologen. Attempts to construct a sodBmutant in this organism by allelic exchange of the chromosomal copy of the gene with a suicide plasmid containing a mutated sodBgene were unsuccessful, strongly suggesting that the encoded superoxide dismutase is essential for viability of R. capsulatus in aerobic cultures.

scholarly journals Identification of Genes of VSH-1, a Prophage-Like Gene Transfer Agent of Brachyspira hyodysenteriae

2005 ◽  
Vol 187 (17) ◽  
pp. 5885-5892 ◽  
Author(s):  
Eric G. Matson ◽  
M. Greg Thompson ◽  
Samuel B. Humphrey ◽  
Richard L. Zuerner ◽  
Thad B. Stanton
Keyword(s):  
Amino Acid ◽  
Gene Transfer ◽  
Rhodobacter Capsulatus ◽  
Bacterial Species ◽  
Polyacrylamide Gel Electrophoresis ◽  
Bacterial Genome ◽  
Sodium Dodecyl ◽  
Amino Acid Sequences ◽  
Brachyspira Hyodysenteriae ◽  
Gene Transfer Agent

ABSTRACT VSH-1 is a mitomycin C-inducible prophage of the anaerobic spirochete Brachyspira hyodysenteriae. Purified VSH-1 virions are noninfectious, contain random 7.5-kb fragments of the bacterial genome, and mediate generalized transduction of B. hyodysenteriae cells. In order to identify and sequence genes of this novel gene transfer agent (GTA), proteins associated either with VSH-1 capsids or with tails were purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequences of 11 proteins were determined. Degenerate PCR primers were designed from the amino acid sequences and used to amplify several VSH-1 genes from B. hyodysenteriae strain B204 DNA. A λ clone library of B. hyodysenteriae B204 DNA was subsequently screened by Southern hybridization methods and used to identify and sequence overlapping DNA inserts containing additional VSH-1 genes. VSH-1 genes spanned 16.3 kb of the B. hyodysenteriae chromosome and were flanked by bacterial genes. VSH-1 identified genes and unidentified, intervening open reading frames were consecutively organized in head (seven genes), tail (seven genes), and lysis (four genes) clusters in the same transcriptional direction. Putative lysis genes encoding endolysin (Lys) and holin proteins were identified from sequence and structural similarities of their translated protein products with GenBank bacteriophage proteins. Recombinant Lys protein hydrolyzed peptidoglycan purified from B. hyodysenteriae cells. The identified VSH-1 genes exceed the DNA capacity of VSH-1 virions and do not encode traditional bacteriophage early functions involved in DNA replication. These genome properties explain the noninfectious nature of VSH-1 virions and further confirm its resemblance to known prophage-like, GTAs of other bacterial species, such as the GTA from Rhodobacter capsulatus. The identification of VSH-1 genes will enable analysis of the regulation of this GTA and should facilitate investigations of VSH-1-like prophages from other Brachyspira species.

Structural analysis of the photosynthetic membrane from Ectothiorhodospira halochloris

1983 ◽  
Vol 41 ◽  
pp. 740-741
Author(s):  
H. Engelhardt ◽  
R. Guckenberger ◽  
W. Baumeister
Keyword(s):  
Lattice Structure ◽  
Bacterial Species ◽  
Hexagonal Lattice ◽  
Reaction Centre ◽  
Photosynthetic Membrane ◽  
Rhodopseudomonas Viridis ◽  
Photosynthetic Membranes ◽  
Ectothiorhodospira Halochloris ◽  
Main Components

Bacterial photosynthetic membranes contain, apart from lipids and electron transport components, reaction centre (RC) and light harvesting (LH) polypeptides as the main components. The RC-LH complexes in Rhodopseudomonas viridis membranes are known since quite seme time to form a hexagonal lattice structure in vivo; hence this membrane attracted the particular attention of electron microscopists. Contrary to previous claims in the literature we found, however, that 2-D periodically organized photosynthetic membranes are not a unique feature of Rhodopseudomonas viridis. At least five bacterial species, all bacteriophyll b - containing, possess membranes with the RC-LH complexes regularly arrayed. All these membranes appear to have a similar lattice structure and fine-morphology. The lattice spacings of the Ectothiorhodospira haloohloris, Ectothiorhodospira abdelmalekii and Rhodopseudomonas viridis membranes are close to 13 nm, those of Thiocapsa pfennigii and Rhodopseudomonas sulfoviridis are slightly smaller (∼12.5 nm).

scholarly journals Development of a Genoserotyping Method for Salmonella Infantis Detection on the Basis of Pangenome Analysis

2020 ◽  
Vol 9 (1) ◽  
pp. 67
Author(s):  
Seung-Min Yang ◽  
Jiwon Baek ◽  
Eiseul Kim ◽  
Hyeon-Be Kim ◽  
Seyoung Ko ◽  
...  
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Cost Effective ◽  
The Other ◽  
Gene Marker ◽  
Diagnostic Assay ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Substantial Economic Burden

In recent years, Salmonella Infantis has become a predominant serovariant in clinical and poultry isolates, thereby imposing a substantial economic burden on both public health and the livestock industry. With the aim of coping with the steep increase in serovar Infantis prevalence, a polymerase chain reaction (PCR)-based rapid and accurate diagnostic assay was developed in this study through pangenome profiling of 60 Salmonella serovars. A gene marker, SIN_02055, was identified, which is present in the S. Infantis genome but not in the pangenome of the other serovars. Primers specific to SIN_02055 were used to accurately detect serovar Infantis, and to successfully differentiate Infantis from the other 59 serovars in real-time PCR with a R2 of 0.999 and an efficiency of 95.76%. The developed method was applied to 54 Salmonella strains belonging to eight dominant serovars, and distinguished Infantis from the other seven serovars with an accuracy of 100%. The diagnostic primer set also did not show false positive amplification with 32 strains from eight non-Salmonella bacterial species. This cost-effective and rapid method can be considered an alternative to the classic serotyping using antisera.

scholarly journals STUDIES ON THE BACTERIOPHAGE OF D'HERELLE

1925 ◽  
Vol 42 (4) ◽  
pp. 483-497 ◽  
Author(s):  
Jacques J. Bronfenbrenner ◽  
Charles Korb
Keyword(s):  
Quantitative Evaluation ◽  
Bacterial Species ◽  
Relative Concentration ◽  
Serial Dilution ◽  
The Other ◽  
Resistant Bacteria ◽  
Dilution Method ◽  
High Concentration ◽  
Average Size ◽  
Broth Dilution

The experiments reported above confirm the fact that lytic principle is distributed in active solution in a state of indivisible units. This permits its quantitative evaluation by serial dilution, as well as by plating on agar. The latter method, however, often gives readings considerably lower than those obtained by the broth dilution method of titration. By varying the concentration of agar it has been possible to show that the discrepancy is due to adsorption of the lytic agent on agar. When the concentration of the latter is increased from 0.3 per cent to 2.5 per cent the number of plaques of lysis is reduced more than 100 times. At the same time the average size of the plaques also decreases approximately to one-tenth of the original. The size, as well as the number of plaques, has been found to depend also on the condition of the culture employed in titration. Thus, when the culture exposed to the action of lytic agent is composed of young susceptible bacteria, the greater the concentration of bacteria, the smaller the plaques. When the culture is composed partly of young and partly of old susceptible bacteria, both the size and the number of the plaques are diminished with the increase in the relative concentration of old bacteria. On the other hand, presence in the culture of resistant bacteria does not affect either the size or the number of the plaques so long as the relative concentration of susceptible bacteria in the culture is sufficient to allow formation of them. The plaques appearing in the presence of a high concentration of resistant variants in the culture are relatively indistinct owing to overgrowth. Under carefully controlled conditions the size of plaques is found to be determined by the character of the lytic filtrate. Thus in the case of lytic agents which act upon more than one bacterial species the size of the plaques remains constant, irrespective of the bacterial substratum used for the production of the active filtrate.

scholarly journals A Method for Fabricating Arrays of Nanopatterns with the Feature Size beyond Diffraction Limit

2008 ◽  
Vol 2008 ◽  
pp. 1-4 ◽  
Author(s):  
Shuhong Li ◽  
Lifang Shi ◽  
Xiaochun Dong ◽  
Chunlei Du ◽  
Yudong Zhang
Keyword(s):  
Time Domain ◽  
Finite Difference Time Domain ◽  
Diffraction Limit ◽  
Evanescent Waves ◽  
The Other ◽  
Optical Intensity ◽  
Polystyrene Spheres ◽  
Feature Size ◽  
Contact Points ◽  
Difference Time

A convenient lithographic technique is proposed in this paper, which can be used to produce subdiffraction-limit arrays of nanopatterns over large areas (about several square centimeters). An array of polystyrene spheres (PS) is arranged on the surface of a layer of silver which has a thickness of about tens of nanometers. With the normal illumination light of wavelength 365 nm perpendicular to the substrate, PS can generate an array of optical patterns with high intensity at their contact points with silver. By designing the silver slab, the evanescent waves that carry subwavelength information about the optical patterns are substantially enhanced, while propagating components are restrained. In the photoresist which is on the other side of silver, the optical intensity is redistributed and subdiffraction-limit patterns are obtained after exposure and development. Simulation by finite-difference time-domain (FDTD) and experiments were carried out to verify the technique. The results show that by using PS with diameter of 600 nm, nanopatterns with dimension of less than 80 nm can be obtained.

scholarly journals Research on reproductive performance of mares in Serbia using bacteriological examination

2011 ◽  
Vol 27 (3) ◽  
pp. 883-892
Author(s):  
M.I. Urosevic ◽  
D. Stojanovic ◽  
B. Lako ◽  
I. Jajic ◽  
Z. Milicic ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Reproductive Performance ◽  
Staphylococcus Epidermidis ◽  
Pasteurella Multocida ◽  
Bacterial Species ◽  
The Other ◽  
Streptococcus Zooepidemicus ◽  
Bacteriological Examination ◽  
Reproductive Disorders ◽  
Bacillus Spp

The research was conducted on 19 stud farms in Serbia, on 80 mares used for breeding, with and without reproductive disorders. During the two years period (from 2009 to 2010) double guarded uterine swabs from 80 mares, aged between 3 and 22 years were collected. Mares belonged to the different breeds: Thoroughbred, Standardbred, Lipizzaner and mixed breeds. It was determined, that bacterial infection of genital organs was found in 24 mares in the examined population, and the bacterial species Streptococcus zooepidemicus was diagnosed in the 11 samples from cervical swabs. In the 5 samples, Escherichia coli was isolated, while Staphylococcus epidermidis and Pasteurella multocida were present in the 2 samples each, while the other causes and simultaneous isolation of two bacterial species are much less present. These species are: Bacillus spp. plus Escherichia coli; Streptococcus zooepidemicus plus Klebsiella pneumoniae and Escherichia coli plus Streptococcus zooepidemicus. In one swab we determinated Arcanobacter pyogenes. In this examination, according to available data after natural mating, we found conception level of 43,10%, which is similar with previous reports in our country.

scholarly journals The Human Cytomegalovirus UL74 Gene Encodes the Third Component of the Glycoprotein H-Glycoprotein L-Containing Envelope Complex

1998 ◽  
Vol 72 (10) ◽  
pp. 8191-8197 ◽  
Author(s):  
Mary T. Huber ◽  
Teresa Compton
Keyword(s):  
Amino Acid ◽  
Human Cytomegalovirus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Open Reading Frames ◽  
Protein Species ◽  
Gene Encoding ◽  
Peptide Backbone ◽  
Glycoprotein H ◽  
Infected Cells

ABSTRACT The human cytomegalovirus (HCMV) gCIII envelope complex is composed of glycoprotein H (gH; gpUL75), glycoprotein L (gL; gpUL115), and a third, 125-kDa protein not related to gH or gL (M. T. Huber and T. Compton, J. Virol. 71:5391–5398, 1997; L. Li, J. A. Nelson, and W. J. Britt, J. Virol. 71:3090–3097, 1997). Glycosidase digestion analysis demonstrated that the 125-kDa protein was a glycoprotein containing ca. 60 kDa of N-linked oligosaccharides on a peptide backbone of 65 kDa or less. Based on these biochemical characteristics, two HCMV open reading frames, UL74 and TRL/IRL12, were identified as candidate genes for the 125-kDa glycoprotein. To identify the gene encoding the 125-kDa glycoprotein, we purified the gCIII complex, separated the components by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and subjected gH and the 125-kDa glycoprotein to amino acid microsequence analysis. Microsequencing of an internal peptide derived from purified 125-kDa glycoprotein yielded the amino acid sequence LYVGPTK. A FASTA search revealed an exact match of this sequence to amino acids 188 to 195 of the predicted product of the candidate gene UL74, which we have designated glycoprotein O (gO). Anti-gO antibodies reacted in immunoblots with a protein species migrating at ca. 100 to 125 kDa in lysates of HCMV-infected cells and with 100- and 125-kDa protein species in purified virions. Anti-gO antibodies also immunoprecipitated the gCIII complex and recognized the 125-kDa glycoprotein component of the gCIII complex. Positional homologs of the UL74 gene were found in other betaherpesviruses, and comparisons of the predicted products of the UL74 homolog genes demonstrated a number of conserved biochemical features.

scholarly journals Proteolytic Processing of Sapovirus ORF1 Polyprotein

2005 ◽  
Vol 79 (12) ◽  
pp. 7283-7290 ◽  
Author(s):  
Tomoichiro Oka ◽  
Kazuhiko Katayama ◽  
Satoko Ogawa ◽  
Grant S. Hansman ◽  
Tsutomu Kageyama ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Proteolytic Processing ◽  
Open Reading Frames ◽  
Rabbit Hemorrhagic Disease Virus ◽  
Translation System ◽  
Hemorrhagic Disease ◽  
Rabbit Hemorrhagic Disease ◽  
Capsid Protein Vp1

ABSTRACT The genome of Sapovirus (SaV), a causative agent of gastroenteritis in humans and swine, contains either two or three open reading frames (ORFs). Functional motifs characteristic to the 2C-like NTPase (NTPase), VPg, 3C-like protease (Pro), 3D-like RNA-dependent RNA polymerase (Pol), and capsid protein (VP1) are encoded in the ORF1 polyprotein, which is afterwards cleaved into the nonstructural and structural proteins. We recently determined the complete genome sequence of a novel human SaV strain, Mc10, which has two ORFs. To investigate the proteolytic cleavage of SaV ORF1 and the function of protease on the cleavage, both full-length and truncated forms of the ORF1 polyprotein either with or without mutation in 1171Cys to Ala of the GDCG motif were expressed in an in vitro coupled transcription-translation system. The translation products were analyzed directly by sodium dodecyl sulfate-polyacrylamide gel electrophoresis or by immunoprecipitation with region-specific antibodies. The ORF1 polyprotein was processed into at least 10 major proteins: p11, p28, p35, p32, p14, p70, p60, p66, p46, and p120. Seven of these products were arranged in the following order: NH2-p11-p28-p35(NTPase)-p32-p14(VPg)-p70(Pro-Pol)-p60(VP1)-COOH. p66, p46 and p120 were precursors of p28-p35 (NTPase), p32-p14 (VPg), and p32-p14 (VPg)-p70 (Pro-Pol), respectively. Mutagenesis in the 3C-like protease motif fully abolished the proteolytic activity. The cleavage map of SaV ORF1 is similar to those of other heretofore known members of the family Caliciviridae, especially to rabbit hemorrhagic disease virus, a member of the genus Lagovirus.

scholarly journals Studies on Platelet Glycoproteins by Surface Labeling and SDS Polyacrylamide Gel Electrophoresis

1977 ◽  
Author(s):  
I. Hagen ◽  
N.O. Solum ◽  
M. Peterka
Keyword(s):  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Conformational State ◽  
Polyacrylamide Gel ◽  
The Other ◽  
Metabolic Inhibitors ◽  
Conventional System ◽  
Platelet Surface ◽  
Release Reaction

Platelet surface (glyco)proteins, and alterations in these in connection with the thrombin-induced release reaction has been studied. Platelets were labeled by lactoperoxidase-catalyzed iodination, and examined by SDS gel electrophoresis in two different gel systems, one conventional(J. Biol. Chem.1969 244 4406), and the other containing urea and EDTA in the gels. In the conventional system the bulk of radioactivity coincided with a PAS band (GP III) of MW about 100, 000. In the other system, the main radioactive peak appeared in the GP II area (MW 120,000), and a shift in the PAS stain intensity from GP III to GP II was seen. Thrombasthenic platelets showed only traces of the GP II band in both systems. The bulk of radioactivity was associated with the surface glycopolypeptide GPS, which is present, but not labeled in normal platelets. In thrombin-released platelets, GPS in its unreduced state has an altered electrophoretic mobility compared to control platelets and platelets which have been incubated with metabolic inhibitors to prevent secretion. The findings indicate that the GP III band consists of two different polypeptides, one of which appears in the GP II area in gels containing urea and EDTA. Further, that thrombasthenic platelet membrane exists in a conformational state different from that of normal platelets. And finally, GPS is in some way involved in, or influenced by, the thrombin-induced release reaction.

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