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1.
Some Bacillus species, especially Bacillus subtilis and Bacillus pumilus groups, have highly similar 16S rRNA gene sequences, which are hard to identify based on 16S rDNA sequence analysis. To conquer this drawback, rpoB, recA sequence analysis along with randomly amplified polymorphic (RAPD) fingerprinting was examined as an alternative method for differentiating Bacillus species. The 16S rRNA, rpoB and recA genes were amplified via a polymerase chain reaction using their specific primers. The resulted PCR amplicons were sequenced, and phylogenetic analysis was employed by MEGA 6 software. Identification based on 16S rRNA gene sequencing was underpinned by rpoB and recA gene sequencing as well as RAPD-PCR technique. Subsequently, concatenation and phylogenetic analysis showed that extent of diversity and similarity were better obtained by rpoB and recA primers, which are also reinforced by RAPD-PCR methods. However, in one case, these approaches failed to identify one isolate, which in combination with the phenotypical method offsets this issue. Overall, RAPD fingerprinting, rpoB and recA along with concatenated genes sequence analysis discriminated closely related Bacillus species, which highlights the significance of the multigenic method in more precisely distinguishing Bacillus strains. This research emphasizes the benefit of RAPD fingerprinting, rpoB and recA sequence analysis superior to 16S rRNA gene sequence analysis for suitable and effective identification of Bacillus species as recommended for probiotic products.  相似文献   

2.
A limitation to the engineering of cellulolytic thermophiles is the availability of functional, thermostable (≥?60 °C) replicating plasmid vectors for rapid expression and testing of genes that provide improved or novel fuel molecule production pathways. A series of plasmid vectors for genetic manipulation of the cellulolytic thermophile Caldicellulosiruptor bescii has recently been extended to Clostridium thermocellum, another cellulolytic thermophile that very efficiently solubilizes plant biomass and produces ethanol. While the C. bescii pBAS2 replicon on these plasmids is thermostable, the use of homologous promoters, signal sequences and genes led to undesired integration into the bacterial chromosome, a result also observed with less thermostable replicating vectors. In an attempt to overcome undesired plasmid integration in C. thermocellum, a deletion of recA was constructed. As expected, C. thermocellum ?recA showed impaired growth in chemically defined medium and an increased susceptibility to UV damage. Interestingly, we also found that recA is required for replication of the C. bescii thermophilic plasmid pBAS2 in C. thermocellum, but it is not required for replication of plasmid pNW33N. In addition, the C. thermocellum recA mutant retained the ability to integrate homologous DNA into the C. thermocellum chromosome. These data indicate that recA can be required for replication of certain plasmids, and that a recA-independent mechanism exists for the integration of homologous DNA into the C. thermocellum chromosome. Understanding thermophilic plasmid replication is not only important for engineering of these cellulolytic thermophiles, but also for developing genetic systems in similar new potentially useful non-model organisms.  相似文献   

3.
Ribosomal protein S1 of Mycobacterium tuberculosis (MtRpsA) binds to ribosome and mRNA, and plays significant role in the regulation of translation initiation, conventional protein synthesis and transfer-messenger RNA (tmRNA) mediated trans-translation. It has been identified as the target of pyrazinoic acid (POA), a bactericidal moiety from hydrolysis of pyrazinamide, which is a mainstay of combination therapy for tuberculosis. POA prevented the interactions between the C-terminal S1 domain of MtRpsA (residues 280–368, MtRpsACTD_S1) and tmRNA; so that POA can inhibit the trans-translation, which is a key component of multiple quality control pathways in bacteria. However, the details of molecular mechanism and dynamic characteristics for MtRpsACTD_S1 interactions with POA, tmRNA or mRNA are still unclear. Here we present the 1H, 15N, 13C resonance assignments of MtRpsACTD_S1 as well as the secondary structure information based on backbone chemical shifts, which lay foundation for further solution structure determination, dynamic properties characterization and interactions investigation between MtRpsACTD_S1 and tmRNA, RNA or POA.  相似文献   

4.
Recombination, synapsis, chromosome segregation and gene expression are co-ordinately regulated during meiosis to ensure successful execution of this specialised cell division. Studies with multiple mutant mouse lines have shown that mouse spermatocytes possess quality control checkpoints that eliminate cells with persistent defects in chromosome synapsis. In addition, studies on Trip13mod/mod mice suggest that pachytene spermatocytes that successfully complete chromosome synapsis can undergo meiotic arrest in response to defects in recombination. Here, we present additional support for a meiotic recombination-dependent checkpoint using a different mutant mouse line, Tex19.1?/?. The appearance of early recombination foci is delayed in Tex19.1?/? spermatocytes during leptotene/zygotene, but some Tex19.1?/? spermatocytes still successfully synapse their chromosomes and we show that these spermatocytes are enriched for early recombination foci. Furthermore, we show that patterns of axis elongation, chromatin modifications and histone H1t expression are also all co-ordinately skewed towards earlier substages of pachytene in these autosomally synapsed Tex19.1?/? spermatocytes. We also show that this skew towards earlier pachytene substages occurs in the absence of elevated spermatocyte death in the population, that spermatocytes with features of early pachytene are present in late stage Tex19.1?/? testis tubules and that the delay in histone H1t expression in response to loss of Tex19.1 does not occur in a Spo11 mutant background. Taken together, these data suggest that a recombination-dependent checkpoint may be able to modulate pachytene progression in mouse spermatocytes to accommodate some types of recombination defect.  相似文献   

5.
An endophytic actinobacterial strain was isolated from a yellowwood tree growing on the slope of Devil’s Peak, Cape Town, South Africa. Analysis of the 16S rRNA gene showed that the strain belongs to the genus Kribbella. Phylogenetic analyses using the 16S rRNA gene and multilocus sequence analysis using the concatenated gene sequences of the gyrB, rpoB, relA, recA and atpD genes showed that strain YPL1T is closely related to the type strains of Kribbella karoonensis and Kribbella shirazensis. DDH experiments showed that strain YPL1T is a distinct genomic species from its close phylogenetic relative, K. karoonensis Q41T. Physiological comparisons further showed that strain YPL1T is phenotypically distinct from the type strains of Kribbella jejuensis, Kribbella aluminosa, K. karoonensis, K. shirazensis and Kribbella swartbergensis. Strain YPL1T is thus presented as the type strain of a novel species, for which the name Kribbella podocarpi sp. nov. (= DSM 29424T = NRRL B-65063T), is proposed.  相似文献   

6.
A novel actinomycete strain designated S2T was isolated from Tunisian rhizosphere soil of Lavandula officinalis. This isolate exhibited broad spectrum antibacterial activity against several Gram-positive and Gram-negative bacteria and also antifungal activity against yeast and filamentous fungi. The isolate S2T presents morphological and chemotaxonomic characteristics typical of the members of the genus Streptomyces. Whole cell hydrolysates of S2T were found to contain LL-diaminopimelic acid. The major fatty acids were identified as C16:0, anteiso-C15:0 and iso-C16:0 whereas the predominant menaquinones were found to be MK-9(H6) and MK-9(H8). The polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and three unidentified compounds. The G+C content of the genomic DNA was determined to be 71.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S2T belongs to the genus Streptomyces and is closely related to Streptomyces netropsis DSM 40259T with 99.86% sequence similarity. Multi-locus sequence analysis (MLSA) based on four house-keeping gene alleles (gyrB, recA, trpB, rpoB) showed that isolate S2T is closely related to S. netropsis, with an MLSA distance greater than 0.007. The DNA–DNA relatedness between strain S2T and its near phylogenetic neighbour was 63.6 ± 2.3%, which is lower than the 70% threshold value for delineation of genomic prokaryotic species. This isolate was also distinguished from the type strain S. netropsis DSM 40259T, using a combination of morphological and physiological features. Based on its phenotypic and molecular properties, strain S2T is considered to represent a novel species of the genus Streptomyces, for which the name Streptomyces tunisialbus sp. nov. is proposed. The type strain is S2T (= JCM 32165T = DSM 105760T).  相似文献   

7.
A pleomorphic Gram-negative, motile coccobacillus was isolated from the gills of a wild-caught bluespotted ribbontail ray after its sudden death during quarantine. Strain 141012304 was observed to grow aerobically, to be clearly positive for cytochrome oxidase, catalase, urease and was initially identified as “Brucella melitensis” or “Ochrobactrum anthropi” by Matrix-assisted laser desorption/ionization-time of flight mass spectrometry and VITEK2-compact®, respectively. Affiliation to the genus Brucella was confirmed by bcsp31 and IS711 PCR as well as by Brucella species-specific multiplex PCR, therein displaying a characteristic banding pattern recently described for Brucella strains obtained from amphibian hosts. Likewise, based on recA sequencing, strain 141012304 was found to form a separate lineage, within the so called ‘atypical’ Brucella, consisting of genetically more distantly related strains. The closest similarity was detected to brucellae, which have recently been isolated from edible bull frogs. Subsequent next generation genome sequencing and phylogenetic analysis confirmed that the ray strain represents a novel Brucella lineage within the atypical group of Brucella and in vicinity to Brucella inopinata and Brucella strain BO2, both isolated from human patients. This is the first report of a natural Brucella infection in a saltwater fish extending the host range of this medically important genus.  相似文献   

8.
A strain 5-1-2T was isolated from a root nodule of Hedysarum multijugum collected from Zhangye city, Gansu province, north-west China. Phylogenetic analysis based on the 16S rRNA gene sequence and other housekeeping genes (recA and atpD) indicated that the strain represents a novel species in the genus Rhizobium close to the strain Rhizobium subbaraonis JC85T with similarities of 98.27, 88.92 and 89.62%, respectively. Strain 5-1-2T contained Q-10 as the predominant ubiquinone. Our results showed that the major fatty acids were feature 8 (C18:1 ω7c and/or C18:1 ω6c; 38.90%). In addition, the DNA–DNA hybridizations with the type strains R. subbaraonis JC85T and Rhizobium halophytocola YC6881T were 39.2 ± 2.1 and 44.3 ± 1.9, respectively. Therefore, a novel species Rhizobium hedysari sp. nov. is proposed, and 5-1-2T (=CGMCC1.15677T = NBRC112532T) is designated as the type strain.  相似文献   

9.
We have devised a system for isolating yeast DNA sequences that are able to act as initiators of recombination leading to deletions in mitotically growing yeast cells. This system has allowed us to identify the FRT site of the 2μ site-specific recombinase Flp as such a sequence. We show that Flp is able to initiate recombination leading to deletions at a single FRT site in cir o strains. These results indicate that Flp is able to cleave a single FRT site, supporting the observation that the mechanism of cleavage by Flp is trans-horizontal. Interestingly, Flp can induce homologous recombination in both a RAD52-dependent and RAD52-independent manner. Our work provides a new system for the study of homologous recombination leading to deletions, in which the initiation step can be efficiently controlled. We discuss the possibility that Flp-induced, RAD52-independent events occur by single-strand annealing.  相似文献   

10.
11.
It is known that the genus Waminoa (order Acoela, Family Convolutidae) associates with anthozoans (Scleractinia, Octocorallia, Zoantharia) and feeds on the mucus of their host. Despite the close relationship between Waminoa and its host species, it is known from phylogenetic analyses of sequences of the internal transcribed spacer 2 of ribosomal DNA (ITS-2) that photosymbiotic Symbiodinium isolated from Waminoa are different from the Symbiodinium of their anthozoan hosts. Additionally, Waminoa inherit their Symbiodinium and Amphidinium via vertical transmission. In the current study, we looked for and collected Waminoa from different anthozoan hosts from the Ryukyu Archipelago in southern Japan, and then examined the Symbiodinium of Waminoa as well as hosts’ Symbiodinium, and utilized sequences of the hyper-variable non-coding region of the plastid minicircle (psbAncr) in order to re-examine the diversity of Symbiodinium within Waminoa. In the resulting psbAncr phylogenetic tree, all Symbiodinium within Waminoa comprised a unique group within Clade C, all with the same genotype in the psbAncr phylogenetic tree. Our results reconfirm that most Waminoa host their own lineage of Symbiodinium, and demonstrate that Waminoa are also found on azooxanthellate anthozoan hosts, as we found Waminoa on Siphonogorgia sp., expanding their potential known habitat. Additionally, two Symbiodinium from Waminoa from both shallow (4 m) and mesophotic (32 m) depths were not included in main group of Symbiodinium from Waminoa in the ITS-2 phylogenetic tree, although we could not acquire psbAncr sequences for these specimens. Therefore it is possible that undetected Symbiodinium diversity may yet exist within Waminoa, and future work at shallow to mesophotic depths examining both zooxanthellate and azooxanthellate hosts may find more Waminoa and Symbiodinium diversity.  相似文献   

12.
The diversity of rhizobia that establish symbiosis with Lotus corniculatus has scarcely been studied. Several species of Mesorhizobium are endosymbionts of this legume, including Mesorhizobium loti, the type species of this genus. We analysed the genetic diversity of strains nodulating Lotus corniculatus in Northwest Spain and ten different RAPD patterns were identified among 22 isolates. The phylogenetic analysis of the 16S rRNA gene showed that the isolated strains belong to four divergent phylogenetic groups within the genus Mesorhizobium. These phylogenetic groups are widely distributed worldwide and the strains nodulate L. corniculatus in several countries of Europe, America and Asia. Three of the groups include the currently described Mesorhizobium species M. loti, M. erdmanii and M. jarvisii which are L. corniculatus endosymbionts. An analysis of the recA and atpD genes showed that our strains belong to several clusters, one of them very closely related to M. jarvisii and the remanining ones phylogenetically divergent from all currently described Mesorhizobium species. Some of these clusters include L. corniculatus nodulating strains isolated in Europe, America and Asia, although the recA and atpD genes have been sequenced in only a few L. corniculatus endosymbionts. The results of this study revealed great phylogenetic diversity of strains nodulating L. corniculatus, allowing us to predict that even more diversity will be discovered as further ecosystems are investigated.  相似文献   

13.
Dermacentor nitens tick is commonly found in the equine auditory canal, where it causes economic losses due to its direct damage, causing blood spoliation, stress, transmission of pathogens, and predisposition to myasis and secondary bacterial infection in its hosts. In this study we evaluated the effect of ethanolic extracts of Cerrado plants on biological parameters of engorged females of D. nitens. Ethanolic extracts were prepared from the leaves of Schinopsis brasiliensis, Piptadenia viridiflora, Ximenia americana, and Serjania lethalis at 25–150 mg mL?1. Groups of 10 engorged adult females were treated with these extracts and compared with a control containing distilled water and another control with organophosphate, using five replicates for each group. Compared with the control with water, S. lethalis and X. americana extracts at 100 and 150 mg mL?1 significantly inhibited the posture ability. Differently, extracts of S. brasiliensis and P. viridiflora were the most effective in inhibiting larval hatching. Extracts of X. americana and P. viridiflora showed effective inhibition of reproductive parameters of the tick, presenting dose-dependent effect with IC90 78.86 and 78.94 mg mL?1, respectively. Theses effective extracts contained low condensed tannin levels and their HPLC chromatograms revealed the presence of flavonoids. The efficacies of P. viridiflora and X. americana extracts were higher than 90% indicating that these extracts are promising as alternative agents for D. nitens control.  相似文献   

14.
The recognition of bacterial functions involved in DNA metabolism of bacteriophage T4 might reveal interactions between different enzymes during DNA replication and recombination. To detect such functions we have studied the replication of complete and incomplete T4 chromosomes in various mutant strains of Escherichia coli that are defective in their own DNA metabolism. We found that several E. coli functions can substitute for phage functions in T4 replication and recombination and will discuss here the role of the E. coli pol A gene which codes for DNA polymerase I1–4 and of the dna B and E genes3,5.  相似文献   

15.
A method for in vivo studying the fidelity of DNA double-strand break (DSB) repair in bacteriophage T4 has been developed. The frequency of reversion of rII mutations to the wild phenotype was measured in i segC + × i ets1 segCΔ crosses, where ets1 is an insertion in the initial part of the rIB gene carrying a sequence recognized by SegC endonuclease; i designates a rIIB or rIIA mutation located at some distance from ets1, and segCΔ is a deletion in the segC gene. In such cross, a DSB occurs in the site of ets1. Their repair involves genetic recombination and DNA replication in the neighborhood of ets1. In parallel, the frequency of reversion of the same i mutant in the absence of DSBs is measured in i × i self-crosses. Reversions of different types (base substitutions, deletions, insertions) can be studied with the use of structurally different i mutations located at varying distances from ets1. The reversion frequencies were determined for three rIIB mutations and one rIIA mutation. The results obtained suggest that DSB repair in bacteriophage T4 is a process of high fidelity with the rate of errors that does not essentially exceed that in the case of usual phage multiplication.  相似文献   

16.
Strains pyc13T and ZGT13 were isolated from Lake Pengyan and Lake Zigetang on Tibetan Plateau, respectively. Both strains were Gram-negative, catalase- and oxidase-positive, aerobic, rod-shaped, nonmotile, and nonflagellated bacteria. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains pyc13T and ZGT13 belong to the genus Halomonas, with Halomonas alkalicola 56-L4-10aEnT as their closest neighbor, showing 97.4% 16S rRNA gene sequence similarity. The predominant respiratory quinone of both strains was Q-9, with Q-8 as a minor component. The major fatty acids of both strains were C18:1ω6c/C18:1ω7c, C16:1ω6c/C16:1ω7c, C16:0, and C12:0 3OH. The polar lipids of both strains consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, glycolipid, phospholipids of unknown structure containing glucosamine, and unidentified phospholipids. The DNA G + C content of pyc13T and ZGT13 were 62.6 and 63.4 mol%, respectively. The DNA-DNA hybridization values of strain pyc13T were 34, 41, 61, 35, and 35% with the reference strains H. alkalicola 56-L4-10aEnT, H. sediminicola CPS11T, H. mongoliensis Z-7009T, H. ventosae Al12T, and H. fontilapidosi 5CRT, respectively. Phenotypic, biochemical, genotypic, and DNA-DNA hybridization data showed that strains pyc13T and ZGT13 represent a new species within the genus Halomonas, for which the name H. tibetensis sp. nov. is proposed. The type strain is pyc13T (= CGMCC 1.15949T = KCTC 52660T).  相似文献   

17.
In this study, we sought to isolate Salmonella Enteritidis-specific lytic bacteriophages (phages), and we found a lytic phage that could lyse not only S. Enteritidis but also other Gramnegative foodborne pathogens. This lytic phage, SS3e, could lyse almost all tested Salmonella enterica serovars as well as other enteric pathogenic bacteria including Escherichia coli, Shigella sonnei, Enterobacter cloacae, and Serratia marcescens. This SS3e phage has an icosahedral head and a long tail, indicating belong to the Siphoviridae. The genome was 40,793 base pairs, containing 58 theoretically determined open reading frames (ORFs). Among the 58 ORFs, ORF49, and ORF25 showed high sequence similarity with tail spike protein and lysozyme-like protein of Salmonella phage SE2, respectively, which are critical proteins recognizing and lysing host bacteria. Unlike SE2 phage whose host restricted to Salmonella enterica serovars Enteritidis and Gallinarum, SS3e showed broader host specificity against Gram-negative enteric bacteria; thus, it could be a promising candidate for the phage utilization against various Gram-negative bacterial infection including foodborne pathogens.  相似文献   

18.
A Gram-stain-negative, facultative aerobic, non-flagellated, and rod-shaped bacterium, designated AR-13T, was isolated from a seawater on the East Sea in South Korea, and subjected to a polyphasic taxonomic study. Strain AR-13T grew optimally at 30°C, at pH 7.0–8.0 and in the presence of 0–0.5% (w/v) NaCl. The phylogenetic trees based on 16S rRNA gene sequences showed that strain AR-13T fell within the clade comprising the type strains of Arcobacter species, clustering coherently with the type strain of Arcobacter venerupis. Strain AR-13T exhibited 16S rRNA gene sequence similarity values of 98.1% to the type strain of A. venerupis and of 93.2–96.9% to the type strains of the other Arcobacter species. Strain AR-13T contained MK-6 as the only menaquinone and summed feature 3 (C16:1ω7c and/or C16:1ω6c), C16:0, C18:1ω7c, and summed feature 2 (iso-C16:1 I and/or C14:0 3-OH) as the major fatty acids. The major polar lipids detected in strain AR-13T were phosphatidylethanolamine, phosphatidylglycerol, and one unidentified aminophospholipid. The DNA G+C content was 28.3 mol% and its mean DNA-DNA relatedness value with the type strain of A. venerupis was 21%. Differential phenotypic properties, together with its phylogenetic and genetic distinctiveness, revealed that strain AR-13T is separated from recognized Arcobacter species. On the basis of the data presented, strain AR-13T is considered to represent a novel species of the genus Arcobacter, for which the name Arcobacter acticola sp. nov. is proposed. The type strain is AR-13T (=KCTC 52212T =NBRC 112272T).  相似文献   

19.
N?-lysine acetylation is a dynamic, reversible, regulatory post-translational modification in prokaryotes and eukaryotes that modulates a variety of protein functions. As known, acetylation is introduced to lysine mainly through two ways in vivo: nonenzymatic acetylation and acetyltransferase/deacetylase system. Herein, we select the Gcn5-like protein acetyltransferase SacAcuA (encoded by SACE_5148) and the sirtuin-type NAD+-dependent deacetylase SacSrtN (encoded by SACE_3798) as the researching objects. By comparison of ΔSACE_3798 and ΔSACE_5148 to wild type of Saccharopolyspora erythraea, the growth and the synthesis of secondary metabolites were affected by SacAcuA/SacSrtN system. Moreover, 96 proteins were classified into three aspects of cellular components, molecular function, and biological processes. These findings suggest that the acetyltransferase/deacetyltransferase system could not only catalyze the acetylation/deacetylation of special proteins but also affect the protein level to modulate the primary and secondary metabolism in S. erythraea.  相似文献   

20.
The main characteristics of a periodic bioleaching of the refractory gold-bearing pyrite ore from the Tandzut deposit (Armenia) with the help of moderate thermophilic bacterium Sulfobacillus thermosulfidooxidans subsp. asporogenes and original thermotolerant strains Leptospirillum spp. were studied. The optimal pH for oxidizing the ore by S. thermosulfidooxidans subsp. asporogenes was 1.8; the pulp density providing maximal iron leaching rate was 10%. The intensity of oxidation processes decreased at higher ore concentrations. When using S. thermosulfidooxidans subsp. asporogenes, the largest amount of iron passed into the solution at the initial oxidant (Fe3+) concentration of 1.3 g/l. Cocultivation of S. thermosulfidooxidans subsp. asporogenes and Leptospirillum spp. increased the degree of pyrite ore leaching to 98.4% vs. 34.1% in the case of the former bacterium alone.  相似文献   

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