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At least 973 records · Page 54Linked to original sources
PubMed1994

Phylogenetic analysis and development of probes for differentiating methylotrophic bacteria.

Fifteen small-subunit rRNAs from methylotrophic bacteria have been sequenced. Comparisons of these sequences with 22 previously published sequences further defined the phylogenetic relationships among these bacteria and illustrated the agreement between phylogeny and physiological characteristics of the bacteria. Phylogenetic trees were constructed with 16S rRNA sequences from methylotrophic bacteria and representative organisms from subdivisions within the class Proteobacteria on the basis of sequence similarities by using a weighted least-mean-square difference method. The methylotrophs have been separated into coherent clusters in which bacteria shared physiological characteristics. The clusters distinguished bacteria which used either the ribulose monophosphate or serine pathway for carbon assimilation. In addition, methanotrophs and methylotrophs which do not utilize methane were found to form distinct clusters within these groups. Five new deoxyoligonucleotide probes were designed, synthesized, labelled with digoxigenin-11-ddUTP, and tested for the ability to hybridize to RNA extracted from the bacteria represented in the unique clusters and for the ability to detect RNAs purified from soils enriched for methanotrophs by exposure to a methane-air atmosphere for one month. The 16S rRNA purified from soil hybridized to the probe which was complementary to sequences present in 16S rRNA from serine pathway methanotrophs and hybridized to a lesser extent with a probe complementary to sequences in 16S rRNAs of ribulose monophosphate pathway methanotrophs. The nonradioactive detection system used performed reliably at amounts of RNA from pure cultures as small as 10 ng.

Base Sequence↗
PubMed2000

Rediscovery of Anopheles (Cellia) clowi (Diptera: Culicidae), a rarely recorded member of the Anopheles punctulatus group.

Anopheline specimens collected in Papua New Guinea were morphologically identified as the rarely recorded Anopheles clowi Rozeboom & Knight. Amplification of the rDNA ITS2 region of this material revealed a fragment of 750 bp confirming its placement in the Anopheles punctulatus group. This group contains 12 species and includes the major malaria vectors in the islands of the southwest Pacific. Digestion of the ITS2 with the restriction enzyme MspI produced restriction fragment-length polymorphism with bands at 380, 300, and 150 bp, a pattern shared by no other members of this group. Phylogenetic analysis involving the sequencing of a 2 kb region of the rDNA 18S gene indicated that An. clowi was monophyletic and basal to the rest of the group and showed considerable independent evolution from the other members. This is the first record of An. clowi in Papua New Guinea and only the third collection of this species since its discovery in 1945.

Animals↗
PubMed1993

Chloroplast transformation in plants: polyethylene glycol (PEG) treatment of protoplasts is an alternative to biolistic delivery systems.

Nicotiana plumbaginifolia protoplasts were directly transformed by PEG treatment with a cloned 16S rRNA gene isolated from a double antibiotic-resistant Nicotiana tabacum plastid mutant. Putative plastid transformants were selected in cell culture by their spectinomycin resistance and identified by their unselected streptomycin resistance. Alternatively, cell lines were selected in the presence of both antibiotics. The cell line (and its regenerated plants) selected solely for spectinomycin resistance demonstrated an extensive segregation of streptomycin resistance in subsequent tests, while the double-selected line showed stable resistance for both antibiotics. The resistance markers were inherited maternally. In the putative plastid transformants the origin of the resistance mutations was identified by the absence of an AatII site, missing in the donor N. tabacum plastid gene (spectinomycin resistance site) but present in that of wild-type N. plumbaginifolia, and a sequence analysis of the particular nucleotide changes in both resistance sites. Restriction enzyme analysis of total plastid DNA (ptDNA), and the recloning and full sequencing of the fragment introduced, investigated in one of the plastid transformants, showed no DNA rearrangements accompanied with the integration process. Sequence analysis indicated a targeted, homologous integration of the DNA fragment introduced but an unexpectedly complete homology of the parental ptDNA sequences in this region prevented the location of borders. Although the frequency of plastid transformant colonies (2 x 10(-5)) should still be improved, this method for stable chloroplast DNA transformation is comparable with or more efficient than the particle bombardment techniques.

Base Sequence↗
PubMed2002

Bioinformatics and high throughput approach to create genomic resources for the study of bovine immunobiology.

The advent of technologies such as real-time reverse transcriptase polymerase chain reaction (RT-PCR) and cDNA microarrays herald a new era in the study of biological systems. In immunobiology, these advances have begun to impact studies of infectious diseases, inflammatory processes, and immune cell function. However, a lack of genetic reagents for domestic and companion animals has precluded widespread application of new technologies to studies in these systems. We have recently described development of cDNA microarrays for studying bovine immunobiology. Although powerful in revealing genes involved in immunological phenomena in cattle, these resources were limited by a lack of genes known to function in immune responses from other species, such as mouse and human. To address this shortcoming, we used a combination of bioinformatics and high throughput RT-PCR to create amplicons representing over 270 bovine genes whose orthologs in other species were known to function in immune responses. Amplified gene segments were prepared from cDNA representing RNA isolated from either unstimulated or concanavalin A (ConA) stimulated peripheral blood mononuclear cells (PBMCs). In total, 276 genes were amplified from cDNA representing unstimulated bovine PBMC RNA or from cDNA representing ConA stimulated bovine PBMC RNA. A web-accessible resource (http://gowhite.ans.msu.edu/public_php/gd-bovine-immunology.php) has been created to assist in dissemination of this novel resource. The web-accessible resource contains information on gene name, the forward and reverse primers used to amplify each segment, expected product size, and if the gene was found in unstimulated PBMCs or only in ConA stimulated PBMCs. Gene names appear as hyperlinks to the Genbank pages representing the bovine gene or expressed sequence tag (EST) used to generate each primer pair.

Animals↗
PubMed1993

Assignment of fatty acid-beta-oxidizing syntrophic bacteria to Syntrophomonadaceae fam. nov. on the basis of 16S rRNA sequence analyses.

After enrichment from Chinese rural anaerobic digestor sludge, anaerobic, sporing and nonsporing, saturated fatty acid-beta-oxidizing syntrophic bacteria were isolated as cocultures with H2- and formate-utilizing Methanospirillum hungatei or Desulfovibrio sp. strain G-11. The syntrophs degraded C4 to C8 saturated fatty acids, including isobutyrate and 2-methylbutyrate. They were adapted to grow on crotonate and were isolated as pure cultures. The crotonate-grown pure cultures alone did not grow on butyrate in either the presence or the absence of some common electron acceptors. However, when they were reconstituted with M. hungatei, growth on butyrate again occurred. In contrast, crotonate-grown Clostridium kluyveri and Clostridium sticklandii, as well as Clostridium sporogenes, failed to grow on butyrate when these organisms were cocultured with M. hungatei. The crotonate-grown pure subcultures of the syntrophs described above were subjected to 16S rRNA sequence analysis. Several previously documented fatty acid-beta-oxidizing syntrophs grown in pure cultures with crotonate were also subjected to comparative sequence analyses. The sequence analyses revealed that the new sporing and nonsporing isolates and other syntrophs that we sequenced, which had either gram-negative or gram-positive cell wall ultrastructure, all belonged to the phylogenetically gram-positive phylum. They were not closely related to any of the previously known subdivisions in the gram-positive phylum with which they were compared, but were closely related to each other, forming a new subdivision in the phylum. We recommend that this group be designated Syntrophomonadaceae fam. nov.; a description is given.

Bacteria, Anaerobic↗
PubMed1993

A weight array method for splicing signal analysis.

A new method of sequence analysis, using a weight array method (WAM), which generalizes the traditional Staden weight matrix method (WMM), is proposed. With the help of a statistical mechanical model, the discriminant function is identified with the energy function describing macromolecular interactions. The method is applied to the study of 5'-splice signals in Schizosaccharomyces pombe pre-mRNA sequences. The results show that there may exist weak pairwise correlations within the signals and that our method can help to better discriminate these signals. Experiments are proposed to test the predictions of the theory.

Animals↗
PubMed1998

Identification of streptococci from Greek Kasseri cheese and description of Streptococcus macedonicus sp. nov.

Taxonomic studies were performed on some Streptococcus-like organisms isolated from naturally fermented Greek Kasseri cheese. By SDS-PAGE analysis of whole-cell proteins the group was found to be quite different from Streptococcus thermophilus. Comparative 16S and 23S rRNA sequence analyses showed that the isolates represent a new species within the genus Streptococcus, where they are most closely related to the Streptococcus bovis cluster. On the basis of these phylogenetic results and some phenotypic differences, a new species, Streptococcus macedonicus, is proposed. The type strain is ACA-DC 206.

Bacterial Proteins↗
PubMed2006

Bacterial community structure, compartmentalization and activity in a microbial fuel cell.

AIMS: To characterize bacterial populations and their activities within a microbial fuel cell (MFC), using cultivation-independent and cultivation approaches. METHODS AND RESULTS: Electron microscopic observations showed that the fuel cell electrode had a microbial biofilm attached to its surface with loosely associated microbial clumps. Bacterial 16S rRNA gene libraries were constructed and analysed from each of four compartments within the fuel cell: the planktonic community; the membrane biofilm; bacterial clumps (BC) and the anode biofilm. Results showed that the bacterial community structure varied significantly between these compartments. It was observed that Gammaproteobacteria phylotypes were present at higher numbers within libraries from the BC and electrode biofilm compared with other parts of the fuel cell. Community structure of the MFC determined by analyses of bacterial 16S rRNA gene libraries and anaerobic cultivation showed excellent agreement with community profiles from denaturing gradient gel electrophoresis (DGGE) analysis. CONCLUSIONS: Members of the family Enterobacteriaceae, such as Klebsiella sp. and Enterobacter sp. and other Gammaproteobacteria with Fe(III)-reducing and electrochemical activity had a significant potential for energy generation in this system. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has shown that electrochemically active bacteria can be enriched using an electrochemical fuel cell.

Anaerobiosis↗
PubMed2006

Characterization of bacterial populations recovered from the teat canals of lactating dairy and beef cattle by 16S rRNA gene sequence analysis.

Bovine mastitis is of major concern to the dairy industry worldwide. The bovine teat canal is the primary route through which pathogens enter the mammary gland. The microflora of the teat canals of dairy and beef cattle was investigated by analysis of 16S rRNA gene sequences. The goal was to examine the global difference between dairy cattle, which are sanitized on a regular basis, and beef cattle, which receive little sanitary management. A diverse population of 16S rRNA gene sequences was recovered from both the dairy and the beef herd samples, with diversity higher in the beef sample. Analysis revealed the presence of 90 operational taxonomic units (OTUs) among 156 sequences, with 45 OTUs in the dairy sample and 53 OTUs in the beef sample. Only eight OTUs were common to both samples. Members of the classes Clostridia and Bacilli dominated both samples, followed by Actinobacteria and Proteobacteria. The dairy sample contained a cluster (20/80 clones) of Staphylococcaceae members, seven of which were identifiable as coagulase-negative Staphylococcus species. The beef sample was dominated by members of the genus Clostridia, many of which have not been previously cultured. The results suggest that the microorganisms present in the bovine teat canal are more diverse than previously described.

Animals↗
PubMed1994

Sequence analysis of rice hoja blanca virus RNA 3.

RNA 3 of rice hoja blanca tenuivirus (RHBV) has 2299 nucleotides and resembles RNA 3 of other tenuiviruses such as maize stripe (MStV) and rice stripe (RStV) viruses in potentially coding with an ambisense strategy for two proteins. Both the viral-sense protein of 23K and the complementary-sense protein of 35K have about 46% amino acid identity with the analogous proteins encoded by RNA 3 of MStV and RStV. As the proteins of MStV and RStV have about 65% identity between themselves, RHBV cannot be a South and Central American strain of the Asian RStV. The intergenic region resembles those of other tenuiviruses, being rich in A and U residues, but its predicted folding pattern is unlike those of other tenuiviruses. Instead, the predicted folding of the intergenic region was indistinguishable from that of the coding regions and there was no evidence for a distinct hairpin-loop structure. The significance to the evolution of tenuiviruses of the similarities that the two proteins have with their analogues in other tenuiviruses is discussed.

Amino Acid Sequence↗
PubMed1993

The phylogeny of the genus Yersinia based on 16S rDNA sequences.

The inter- and intrageneric relationships of the genus Yersinia were investigated by sequence analysis of the 16S rRNA gene. A stretch of approximately 1450 nucleotides was sequenced from representatives of ten of the eleven validly described species. Phylogenetic analysis revealed that yersinae form a coherent cluster within the gamma subgroup of Proteobacteria. The intrageneric relationship was characterized by five sublines with Y. enterocolitica, Y. rohdei, and Y. ruckeri forming separate sublines each represented by a single species. A separate subline was formed by Y. pestis, Y pseudotuberculosis and Y. kristensenii, while Y. mollaretii, Y. intermedia, Y. bercovieri, Y. aldovae, and Y. kristensenii formed a fifth subline. The phylogenetic distinctness of the yersiniae sublines is compared to published phenotypic properties and results of DNA-DNA similarity studies.

Base Sequence↗
PubMed2001

Longitudinal monitoring of the dynamics of infections due to Bartonella species in UK woodland rodents.

Blood samples were repeatedly collected from 12 sympatric woodland rodents over a 12-month period and DNA extracts from each were incorporated into a bartonella-specific PCR targeting a fragment of the 16S/23S rRNA intergenic spacer region (ISR). The composition of each amplicon was analysed using restriction enzyme analysis (REA) and base sequence comparison. Bartonella DNA was detected in 70 of 109 samples. Eleven samples contained DNA derived from more than one strain. Sequence analysis of 62 samples found 12 sequence variants (ISR genotypes) that were provisionally assigned to 5 different species, 2 of which were newly recognized. Up to five different species were detected in each animal. On about two-thirds of occasions, a species detected I month was not there the next, but never was a genotype superseded by another of the same species. However, a genotype could be re-encountered months later in the same animal, even if interim samples contained other genotypes. Our results suggest that although most animals are bacteraemic most of the time, specific infections are often superseded and that a complex and dynamic epidemiology of bartonella bacteraemias exists in woodland rodents.

Animals↗
PubMed2001

Phenotypic and genotypic characterization of esterase-producing Ureibacillus thermosphaericus isolated from an aerobic digestor of swine waste.

Eight closely related thermophilic strains were isolated from an aerobic and thermophilic treatment of swine wastes. The pleomorphic cells (short and long rods; cocci) showed peritrichous flagella, terminally swollen sporangium, and liberated spores exhibiting hairy appendages. The Gram reaction was negative for both young (4 h) and old (48 h) cultures. Several features, such as colonial morphology, growth between 35 degrees C and 65 degrees C, presence of catalase, presence of spores, and strictly aerobic metabolism (except for one strain), are similar to those found for the genus Bacillus. The inability of the strains to use sugars, except esculin, as source of carbon and energy and the whole cell fatty acid composition are similar to those found in Bacillus thermosphaericus DSM 10633. Sequence analysis of the 16S rRNA gene revealed 99.8%-99.9% identity for seven of the thermophilic strains with this species. A new genus, Ureibacillus, was recently proposed for type strain B. thermosphaericus DSM 10633 The last strain exhibits 97.8% and 97.3% identity with Ureibacillus terrenus DSM12654 and Bacillus sp. TP-84, respectively. Esterase activities were detected for all strains, and assays on p-nitrophenyl butyrate and p-nitrophenyl caprylate revealed that strains were more active on the shorter substrate.

Aerobiosis↗
PubMed2004

Identification of Corynebacterium bovis by endonuclease restriction analysis of the 16S rRNA gene sequence.

Despite its high prevalence within the bovine mammary gland, Corynebacterium bovis is considered a minor pathogen and of limited clinical significance. It has been suggested that intramammary infection with C. bovis may protect quarters against subsequent infection with other pathogens. The literature has produced much conflicting data on the subject. A possible explanation for some of the divergence of opinion on the subject is incorrect identification of isolates in previous studies. This paper describes a novel method for differentiating C. bovis from other lipophilic Corynebacterium species based on endonuclease restriction analysis. The 16S rRNA gene sequences for all known lipophilic Corynebacterium species were obtained from published data and analyzed. It was predicted that endonuclease restriction with HindIII and SmaI could be used to differentiate C. bovis from all other known lipophilic Corynebacterium species. The method was successfully employed to identify 741 of 762 (97.2%) lipophilic Corynebacterium species as C. bovis. Twenty one (2.8%) were identified as species other than C. bovis. Using this technique, it was demonstrated that it is not safe to assume that all lipophilic coryneform organisms isolated from bovine milk samples are C. bovis. This method is an alternative to more traditional methods of identification in large scale studies until methods such as 16S rRNA gene sequencing become more widely available.

Animals↗
PubMed2002

Notes on coccidian phylogeny, based on the apicoplast small subunit ribosomal DNA.

We performed a phylogenetic analysis, based on the partial small subunit rRNA gene (SSU rRNA) sequences from 13 apicoplasts (including new sequences of Sarcocystis muris and Hyaloklossia lieberkuehni) and 16 other plastids, with cyanobacteria as an outgroup. The apicoplast sequences formed a highly supported monophyletic clade with two distinct clades, representing coccidia and haemosporidia, with coccidia divided into Eimeriidae and Sarcocystidae subclades. We confirmed the phylogenetic position of H. lieberkuehni within the Sarcocistidae, as a sister to the Toxoplasma/Neospora group. The coccidian plastid sequences appear to evolve slowly, while their homologues from haemosporidians are more rapidly evolving. We suggest that the higher evolutionary rate is reflected by the increase in the AT content and the possible reduction of the outer apicoplast membrane in some haemosporidians. Since the apicoplast SSU rRNA gene sequences, when compared with their nuclear homologues, offer a higher number of informative positions, they can be used for phylogenetic inference within the Apicomplexa.

Animals↗
PubMed2004

Structural variant of the intergenic internal ribosome entry site elements in dicistroviruses and computational search for their counterparts.

The intergenic region (IGR) located upstream of the capsid protein gene in dicistroviruses contains an internal ribosome entry site (IRES). Translation initiation mediated by the IRES does not require initiator methionine tRNA. Comparison of the IGRs among dicistroviruses suggested that Taura syndrome virus (TSV) and acute bee paralysis virus have an extra side stem loop in the predicted IRES. We examined whether the side stem is responsible for translation activity mediated by the IGR using constructs with compensatory mutations. In vitro translation analysis showed that TSV has an IGR-IRES that is structurally distinct from those previously described. Because IGR-IRES elements determine the translation initiation site by virtue of their own tertiary structure formation, the discovery of this initiation mechanism suggests the possibility that eukaryotic mRNAs might have more extensive coding regions than previously predicted. To test this hypothesis, we searched full-length cDNA databases and whole genome sequences of eukaryotes using the pattern matching program, Scan For Matches, with parameters that can extract sequences containing secondary structure elements resembling those of IGR-IRES. Our search yielded several sequences, but their predicted secondary structures were suggested to be unstable in comparison to those of dicistroviruses. These results suggest that RNAs structurally similar to dicistroviruses are not common. If some eukaryotic mRNAs are translated independently of an initiator methionine tRNA, their structures are likely to be significantly distinct from those of dicistroviruses.

Computational Biology↗
PubMed1995

Primary structure and sequence analysis of RNA2 of a mechanically transmitted barley mild mosaic virus isolate: an evolutionary relationship between bymo- and furoviruses.

The RNA2 nucleotide sequence of a mechanically transmitted isolate of barley mild mosaic virus (BaMMV) has been determined. A combination of Northern blot and sequence analysis indicates that this RNA2 lacks approximately 1000 nucleotides of its C-terminal protein (P2) gene, with respect to Polymyxa graminis transmitted BaMMV. This is confirmed by sequence comparison with RNA2 of a fungus transmitted BaMMV isolate, which reveals the presence of a single deletion located in the 3'-terminal part of the P2 gene. As a consequence, this RNA2 codes for a P2 protein of only 34 K. Sequence homology between the bymovirus P2 protein and the capsid readthrough protein of beet necrotic yellow vein virus and soil-borne wheat mosaic virus suggests an evolutionary relationship between bymo- and furoviruses.

Amino Acid Sequence↗
PubMed1993

Mycobacterium intermedium sp. nov.

Strains of a new type of slowly growing mycobacterium were repeatedly isolated from sputum from a patient with pulmonary disease. This photochromogenic organism grew at 22, 31, 37, and 41 degrees C, possessed catalase, acid phosphatase, esterase, beta-galactosidase, and arylsulfatase activities, and hydrolyzed Tween. It did not produce nicotinic acid or have nitrate reductase, acetamidase, benzamidase, isonicotinamidase, nicotinamidase, pyrazinamidase, succinidamidase, and acid phosphatase activities. Urease activity was variable. The organism is susceptible to ethambutol and resistant to isoniazid and streptomycin. A mycolic acid analysis revealed the presence of alpha-mycolates, alpha'-mycolates, and keto-mycolates. The results of comparative 16S rRNA sequencing placed this organism at an intermediate position between the rapidly and slowly growing mycobacteria. On the basis of the pattern of enzymatic activities and metabolic properties, the results of fatty acid analyses, and the unique 16S rRNA sequence, we propose that this organism represents a new species, for which we propose the name Mycobacterium intermedium. The type strain is strain 1669/91; a culture of this strain has been deposited in the Deutsche Sammlung von Mikroorganismen und Zellkulturen as strain DSM 44049.

Bacterial Typing Techniques↗