ABSTRACT
A polyphasic taxonomic characterization of two novel strain pairs (designated zg-579T/zg-578 and zg-536T/zg-ZUI104) isolated from the faeces of Marmota himalayana was conducted based on phylogenetic analysis of the nearly full-length 16S rRNA gene and genome, digital DNA-DNA hybridization, ortho-average nucleotide identity (Ortho-ANI), and phenotypic and chemotaxonomic traits. Comparative analysis of the nearly full-length 16S rRNA gene sequences showed that strain zg-579T was most closely related to Nocardioides dokdonensis FR1436T (97.57â%) and Nocardioides deserti SC8A-24T (97.36â%), whereas strain zg-536T had the highest similarity to Nocardioides caeni MN8T (98.33â%), Nocardioides convexus W2-2-3T (98.26â%) and Nocardioides daeguensis 2C1-5T (98.19â%). Low levels of DNA-DNA relatedness and Ortho-ANI values (19.8-31.0â%/78.6-88.2â%, zg-579T; 19.9-31.3â%/78.8-86.2â%, zg-536T) between the two new type strains and previously known species within the genus Nocardioides support the hypothesis that the four newly characterized strains could be considered to represent two novel species within this genus. The dominant cellular fatty acids found in strain pair zg-536T/zg-ZUI104 were iso-C16â:â0 and C18â:â1 ω9c, whereas C17â:â1 ω8c was major component in zg-579T/zg-578. Galactose and ribose were the main cell-wall sugars in these two new strain pairs. Diphosphatidylglycerol (DPG), phosphatidylcholine, phosphatidylglycerol (PG) and phosphatidylinositol (PI) were the major polar lipids in zg-579T, whereas DPG, PG and PI predominated in zg-536T. Both strain pairs had MK8(H4) as the major respiratory quinone and ll-diaminopimelic acid as the major cell-wall peptidoglycan. The optimal growth conditions for the two novel strain pairs were 30 °C, pH 7.0 and 0.5â% NaCl (w/v). Based on these polyphasic characterizations, two novel species within the genus Nocardioides are proposed, i.e. Nocardioides marmotae sp. nov. and Nocardioides faecalis sp. nov., with zg-579T (=CGMCC 4.7663T=JCM 33892T) and zg-536T (=CGMCC 4.7662T=JCM 33891T) as the type strains.
Subject(s)
Actinomycetales , Fatty Acids , Fatty Acids/chemistry , Phospholipids/chemistry , Nocardioides , Phylogeny , RNA, Ribosomal, 16S/genetics , DNA, Bacterial/genetics , Base Composition , Sequence Analysis, DNA , Bacterial Typing Techniques , CardiolipinsABSTRACT
Four obligatory anaerobic, Gram-stain-positive, non-motile and rod-shaped organisms (HF-1365T, HF-1362, HF-1101T and HF-4214) were isolated from faecal samples of healthy Chinese subjects. Results of 16S rRNA gene sequence analyses showed that these isolates belong to the genera Enorma (strains HF-1365T and HF-1362) and Eggerthella (strains HF-1101T and HF-4214), closest to Enorma massiliensis (both 98.6â%) and Eggerthella sinensis (98.0 and 97.8â%), respectively. The whole genome sequences of strains HF-1365T and HF-1101T were 2.3 and 4.2 Mb in size with 61.7 and 66.2 mol% DNA G+C content, respectively. The average nucleotide identity and digital DNA-DNA hybridization values indicated that strains HF-1365T and HF-1101T represent novel species in the genera Enorma and Eggerthella. Major fatty acid constituents (>10â%) of strains HF-1365T and HF-1362 were C12â:â0 (24.7 and 23.9â%), C14â:â0 (21.9 and 20.6â%) and summed feature 1 (C15â:â1iso H/C13â:â0 3OH; 12.8 and 10.8â%); those of strains HF-1101T and HF-4214 were C18â:â1 ω9c (32.4 and 33.1â%) and C16â:â0 (13.9 and 14.0â%). Strain HF-1365T had phospholipid, glycolipid, lipid and phosphoglycolipid without any known quinones, while strain HF-1101T had diphosphatidylglycerol as the major polar lipid and MK-7 (80.7â%) as the predominant quinone. On the basis of their phylogenetic and phenotypic characteristics, strains HF-1365T and HF-1101T represent two distinct species, respectively, in the genera Enorma and Eggerthella, for which the names Enorma shizhengliae sp. nov. (type strain HF-1365T=CGMCC 1.17435T=GDMCC 1.1705T=JCM 33601T) and Eggerthella guodeyinii sp. nov. (type strain HF-1101T=CGMCC 1.17436T=GDMCC 1.1668T=JCM 33773T) are proposed.
Subject(s)
Actinobacteria/classification , Feces/microbiology , Phylogeny , Actinobacteria/isolation & purification , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Humans , Nucleic Acid Hybridization , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistryABSTRACT
Four unknown strains, characterized as Gram-stain-negative, strictly anaerobic, non-motile and rod-shaped, were isolated from fresh faeces of healthy humans in PR China. Pairwise sequence comparisons of the 16S rRNA genes showed that these isolates were separated into two clusters. Cluster I (strains HF-5141T and HF-106) was most closely related to Bacteroides xylanisolvens XB1AT (98.0-98.3â% similarity) and Bacteroides ovatus ATCC 8483T (97.3-97.5â%), whereas cluster II (strains HF-5287T and HF-5300) exhibited a similarity range of 96.8-97.0â% to Bacteroides finegoldii JCM 13345T, 96.7-96.9â% to Bacteroides faecis MAJ27T and 96.4-96.5â% to Bacteroides xylanisolvens XB1AT. The DNA G+C contents of type strains HF-5141T and HF-5287T were 41.5 and 42.6 mol%, respectively. These strains had anteiso-C15â:â0 as the major cellular fatty acid, MK-9 and MK-11 as the predominant respiratory quinones, and phosphatidylethanolamine, aminophospholipids and phospholipids as major polar lipids, which is typical for members of the genus Bacteroides. However, the average nucleotide identity and digital DNA-DNA hybridization values, accompanied by different phenotypic and biochemical characteristics, distinguished them from their corresponding closest relatives as well as from other recognized members of the genus Bacteroides. Therefore, strains HF-5141T and HF-5287T represent two novel species in the genus Bacteroides, for which the names Bacteroides luhongzhouii sp. nov. and Bacteroides zhangwenhongii sp. nov. are proposed, with HF-5141T (=CGMCC 1.16787T=GDMCC 1.1591T=JCM 33480T) and HF-5287T (=CGMCC 1.16724T=GDMCC 1.1590T=JCM 33481T) as type strains.
Subject(s)
Bacteroides/classification , Feces/microbiology , Phylogeny , Bacterial Typing Techniques , Bacteroides/isolation & purification , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Humans , Nucleic Acid Hybridization , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Vitamin K 2/chemistryABSTRACT
Six novel facultatively anaerobic, Gram-stain-positive, rod-shaped, non-haemolytic bacteria (zg-320T/zg-336, zg-917T/zg-910 and zg-913T/zg-915) isolated from animal tissues and human faeces were found to belong to the genus Corynebacterium based on the phylogenetic analyses of 16S rRNA gene and 262 core genes set. Based on the greatest degree of 16S rRNA similarity, zg-320T/zg-336 had the highest 16S rRNA gene similarity to Corynebacterium falsenii DSM 44353T (97.51â%), zg-917T/zg-910 to Corynebacterium coyleae DSM 44184T (98.68â%), and zg-913T/zg-915 to Corynebacterium afermentans subsp. lipophilum CIP 103500T (98.79â%). The three novel type strains had a relatively high DNA G+C content (61.2-64.4âmol%), low DNA relatedness and ANI values with their respective neighbours: 23.5/72.7â%, 25.0/72.3%and 22.6/73.1â% (zg-320T vs. Corynebacterium auriscanis CIP 106629T, Corynebacterium resistens DSM 45100T and Corynebacterium suicordis DSM 45110T); 24.4/82.3% and 23.7/81.3â% (zg-917T vs. C. coyleae DSM 44184T and Corynebacterium jeddahense JCBT); 26.8/83.7% and 27.7/84.4â% (zg-913T vs. Corynebacterium mucifaciens ATCC 700355T and C. afermentans subsp. lipophilum CCUG 32105T). The three novel species had C16â:â0, C18â:â0, C18â:â1 ω9c and C18â:â0 ante/C18â:â2 ω6,9c as the major cellular fatty acids; MK-8(H2) in strain zg-917T and MK-9(H2) in strains zg-320T and zg-913T were found to be the major respiratory quinones. For the three novel species, the detected major polar lipids included diphosphatidylglycerol, phosphatidyl inositol mannoside, phosphatidylglycerol and phosphatidylinositol, the cell-wall peptidoglycan was based on meso-DAP, and the whole-cell sugars mainly included ribose, arabinose and galactose. The three novel species grew optimally at 35-37 °C, 0.5â% (w/v) NaCl and pH 7.0-8.0; notably, they were tolerant of 10.5â% (w/v) NaCl. Based on the results of these comprehensive analyses, three novel species in the genus Corynebacterium are proposed, aptly named Corynebacterium zhongnanshanii sp. nov. (zg-320T = GDMCC 1.1719T = JCM 34106T), Corynebacterium lujinxingii sp. nov. (zg-917T = GDMCC 1.1707T = JCM 34094T) and Corynebacterium wankanglinii sp. nov. (zg-913T = GDMCC 1.1706T = JCM 34398T).
Subject(s)
Corynebacterium/classification , Feces/microbiology , Marmota , Phylogeny , Trachea , Animals , Bacterial Typing Techniques , Base Composition , Corynebacterium/isolation & purification , DNA, Bacterial/genetics , Fatty Acids/chemistry , Humans , Marmota/microbiology , Nucleic Acid Hybridization , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Trachea/microbiology , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistryABSTRACT
Four novel strains isolated from the cloacal contents of snow finches (Montifringilla taczanowskii) were characterized as aerobic, Gram-stain-negative, slightly motile, and rod-shaped. Analysis of the 16S rRNA gene sequence revealed that strain CF-458T had the highest similarities of 96.9 and 96.4â% with Limnobaculum parvum HYN0051T and Pragia fontium DSM 5563T, while strain CF-1111T shared the highest similarities of 96.4 and 96.1â% with Pantoea rodasii LMG 26273T and Pectobacterium punjabense SS95T. Phylogenomic analysis showed the four isolates were separated into group â (CF-458T and CF-917) and group â ¡ (CF-1111T and CF-509), and clustered independently in the vicinity of the genera Limnobaculum and Pragia. Summed feature 3 (C16â:â1 ω7c and/or C16â:â1 ω6c, 23.9 and 17.2â%, respectively), C16â:â0 (21.8 and 22.1â%, respectively) and C14â:â0 (10.6 and 17.7â%, respectively) were the common major fatty acids, and summed feature 8 (C18â:â1 ω7c and/or C18â:â1 ω6c, 12.3â%) was also a major fatty acid for strain CF-458T while cyclo-C17â:â0 (13.1%) was for strain CF-1111T. Both had Q-8 as the sole quinone and contained phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol as the major polar lipids. The DNA G+C content of strains CF-458T and CF-1111T was 45.7 and 45.4 mol%, respectively. Based on taxonomic position in the phylogenomic tree and phenotypic properties, two novel species of a new genus within the family Budviciaceae are thus proposed, with the name Jinshanibacter gen. nov., zhutongyuii sp. nov. (type strain CF-458T=CGMCC 1.16483T=GDMCC 1.1586T=JCM 33489T) and Jinshanibacter xujianqingii sp. nov. (type strain CF-1111T=CGMCC 1.16786T=GDMCC 1.1587T=JCM 33490T), respectively.
Subject(s)
Cloaca/microbiology , Finches/microbiology , Gammaproteobacteria/classification , Phylogeny , Animals , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Gammaproteobacteria/isolation & purification , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/chemistryABSTRACT
A Gram-stain-positive, coccus-shaped, non-motile bacterium, designated CF-49T, was isolated from the cloacal content of a snow finch, which was incidentally captured in a plateau pika burrow on the Qinghai-Tibet Plateau, PR China. Analysis of the 16S rRNA gene sequence showed that strain CF-49T was closely related to Vagococcus elongatus CCUG 51432T (96.5â% similarity), Vagococcus fluvialis NCFB 2497T (96.0â%) and Vagococcus lutrae CCUG 39187T (95.9â%), whereas the similarity to another isolate (CF-210) was 99.9â%. Strains CF-49T and CF-210 grew optimally at 37 °C and pH 7.0 and in the presence of 0.5â% (w/v) NaCl. Acid was produced from N-acetylglucosamine, cellobiose, d-fructose, d-glucose, d-mannose, d-mannitol, maltose, d-ribose and salicin. The cell-wall peptidoglycan type was A4α (l-Lys-d-Asp). The major cellular fatty acids (>10â%) were C16â:â0 (35.6â%), C14â:â0 (17.3â%), C18â:â1 ω9c (16.2â%) and C16â:â1 ω9c (10.6â%). The predominant respiratory quinone was menaquinone MK-7 (68.8â%). The G+C content of the genomic DNA was 35.9 mol%. Digital DNA-DNA hybridization of strain CF-49T with V. fluvialis DSM 5731T, V. elongatus CCUG 51432Tand V. lutrae CCUG 39187T resulted in relatedness values of 21.4, 23.3 and 24.6â%, respectively. Based on results from polyphasic analyses, our two isolates are proposed to represent a novel species in the genus Vagococcus, with the name Vagococcus xieshaowenii. The type strain is CF-49T (=CGMCC 1.6436T=GDMCC 1.1588T=JCM 33477T).
Subject(s)
Cloaca/microbiology , Enterococcaceae/classification , Finches/microbiology , Phylogeny , Animals , Bacterial Typing Techniques , Base Composition , Cell Wall/chemistry , DNA, Bacterial/genetics , Enterococcaceae/isolation & purification , Fatty Acids/chemistry , Nucleic Acid Hybridization , Peptidoglycan/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Tibet , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistryABSTRACT
Four unknown strains belonging to the genus Arthrobacter were isolated from plateau wildlife on the Qinghai-Tibet Plateau of PR China. Phylogenetic analysis based on 16S rRNA gene sequences showed that the four isolates were separated into two clusters. Cluster I (strains 785T and 208) had the greatest 16S rRNA gene sequence similarity to Arthrobacter citreus (98.6 and 98.7â%, respectively), Arthrobacter luteolus (98.0 and 98.1%, respectively), Arthrobacter gandavensis (97.9 and 98.0â%, respectively) and Arthrobacter koreensis (97.6 and 97.7â%, respectively). Likewise, cluster II (strains J391T and J915) had the highest sequence similarity to Arthrobacter ruber (98.6 and 98.3â%, respectively) and Arthrobacter agilis (98.1 and 97.9ââ%, respectively). Average nucleotide identity and the digital DNA-DNA hybridization values illustrated that the two type strains, 785T and J391T, represented two separate novel species that are distinct from all currently recognized species in the genus Arthrobacter. These strains had DNA G+C contents of 66.0-66.1 mol% (cluster I) and 68.0 mol% (cluster II). The chemotaxonomic properties of strains 785T and J391T were in line with those of the genus Arthrobacter: anteiso-C15:0 (79.3 and 40.8â%, respectively) as the major cellular fatty acid, MK-8(H2) (65.8â%) or MK-9(H2) (75.6â%) as the predominant respiratory quinone, a polar lipid profile comprising diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, glycolipids and phospholipid, and A3α or A4α as the cell wall peptidoglycan type. On the basis of our results, two novel species in the genus Arthrobacter are proposed, namely Arthrobacter yangruifuii sp. nov. (type strain, 785T=CGMCC 1.16725T=GDMCC 1.1592T=JCM 33491T) and Arthrobacter zhaoguopingii sp. nov. (type strain, J391T=CGMCC 1.17382T=GDMCC 1.1667T=JCM 33841T).
Subject(s)
Arthrobacter/classification , Feces/microbiology , Phylogeny , Animals , Arthrobacter/isolation & purification , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Ducks/microbiology , Equidae/microbiology , Fatty Acids/chemistry , Lagomorpha/microbiology , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistryABSTRACT
Two novel Gram-stain-positive, irregular rod-shaped bacterial strains, dk3136T and dk3543, were isolated from the faeces of Tibetan gazelle (Procapra picticaudata) in the Qinghai-Tibet Plateau of PR China. The cells were aerobic, oxidase-negative and catalase-positive. Colonies were yellowish, circular without any observable aerial mycelium after culturing at 28 â for 3 days on brain-heart infusion (BHI) agar with 5â% sheep blood. The cells grew optimally at 28 °C, pH 7.5 and with 1â% (w/v) NaCl on BHI agar supplemented with 5â% sheep blood. Phylogenetic analysis of the 16S rRNA gene sequences revealed that their nearest phylogenetic relative was Nocardioides solisilvae Ka25T (97.9â% similarity). The results of 16S rRNA gene sequence and phylogenetic/phylogenomic analyses illustrated that N. solisilvae Ka25T, Nocardioides gilvus XZ17T, Nocardioides houyundeii 78T and Nocardioides daphniae D287T were their nearest phylogenetic neighbours. The DNA G+C contents of strains dk3136T and dk3543 were 70.3 mol% and 70.4 mol%, respectively. Their genomes exhibit lower than threshold (95-96â%) average nucleotide identity to known species of the genus Nocardioides. ll-2,6-diaminopimelic acid was the diagnostic diamino acid and MK-8(H4) was the predominant respiratory quinone. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The two strains had C18â:â1 ω9c, iso-C16â:â0 and C17â:â1 ω8c as the major fatty acids, and rhamnose and galactose as the main whole-cell sugars. On the basis of the results of our genotypic, phenotypic and biochemical analyses, we conclude that strains dk3136T and dk3543 represent a novel species in genus Nocardioides, for which the name Nocardioides jishulii sp. nov. is proposed. The type strain is dk3136T (=CGMCC 4.7570T=JCM 33496T=KCTC 49314T).
Subject(s)
Actinobacteria/classification , Antelopes/microbiology , Feces/microbiology , Phylogeny , Actinobacteria/isolation & purification , Animals , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Peptidoglycan/chemistry , Phospholipids/chemistry , Pigmentation , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Tibet , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistryABSTRACT
Two Gram-stain-positive, irregular rod-shaped (0.2-0.5×1.3-2.5 um) strains, HY056T and HY057, were isolated from the faeces of Equus kiang (the largest of the wild asses) collected at different regions from the Qinghai-Tibetan Plateau of PR China. Phylogenetic analyses indicated that strains HY056T and HY057 belong to the genus Nocardioides by sharing a similarity ranging from 96.3 to 97.0â% in the 16S rRNA gene sequence and forming a distinct cluster with Nocardioides daphniae JCM 16608T, Nocardioides houyundeii 78T, Nocardioides solisilvae Ka25T and Nocardioides gilvus XZ17T. The digital DNA-DNA hybridization value of strain HY056T was 96.9â% with strain HY057, but less than 30.0â% with the above four closest relatives. MK-8(H4) was the predominant (91.6â%) respiratory quinone. The cell wall contained ll-2,6-diaminopimelic acid as the diamino acid of the peptidoglycan. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phospholipid. The predominant fatty acids (>10.0â%) were C18â:â1ω9c and iso-C16:0. The DNA G+C contents of strains HY056T and HY057 were 68.9 and 69.1 mol%, respectively. ß-Glucosidase expression was positive, and acid was produced from d-fructose. Strain HY056T (=CGMCC 4.7563T=JCM 33399T) is assigned as the type strain of a novel species within the genus Nocardioides, for which the name Nocardioidesyefusunii sp. nov is proposed.
Subject(s)
Actinobacteria/classification , Equidae/microbiology , Feces/microbiology , Phylogeny , Actinobacteria/isolation & purification , Animals , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Diaminopimelic Acid/chemistry , Fatty Acids/chemistry , Nucleic Acid Hybridization , Peptidoglycan/chemistry , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Tibet , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistryABSTRACT
A cascade six-level phase-mask achromatic coronagraph is proposed. We show that, for the ideal aberration-free case, the average suppression of the central starlight in a three-stage six-level phase-mask achromatic coronagraph (SLPMC) can reach about 10â»8 over a 40% bandwidth. We also do the optimization analysis of the working wavelength of each phase mask in the three-stage SLPMC. With optimization, the starlight can be stably suppressed to less than 10â»8 (average about 10⻹°) over the whole 40% bandwidth.
ABSTRACT
The species in the genus Erwinia are Gram-stain-negative, facultatively anaerobic, motile, and rod-shaped. Most species in the genus Erwinia are phytopathogens. Also, Erwinia persicina was involved in several human infections. Based on the reverse microbial etiology principles, it is worth analyzing the pathogenicity of species in this genus. In this study, we isolated and sequenced two species of Erwinia. Phylogenetic, phenotypic, biochemical, and chemotaxonomic analyses were performed to identify its taxonomy position. The virulence tests on plant leaves and pear fruits were used to identify the plant pathogenicity of two species of Erwinia. Bioinformatic methods predicted the possible pathogenic determinants based on the genome sequence. Meanwhile, adhesion, invasion, and cytotoxicity assays on RAW 264.7 cells were applied to identify animal pathogenicity. We isolated two Gram-stain-negative, facultatively anaerobic, motile, and rod-shaped strains from the feces of ruddy shelducks in the Tibet Plateau of China, designated J780T and J316. Distinct phylogenetic, genomic, phenotypic, biochemical, and chemotaxonomic characters of J780T and J316 identified they were novel species and belonged to the genus Erwinia, for which the name Erwinia sorbitola sp. nov. was proposed, the type strain was J780T (= CGMCC 1.17334T = GDMCC 1.1666T = JCM 33839T). Virulence tests showed blight and rot on the leaves and pear fruits confirmed Erwinia sorbitola sp. nov. was a phytopathogen. Predicted gene clusters of motility, biofilm formation, exopolysaccharides, stress survival, siderophores, and Type VI secretion system might be the causes of pathogenicity. In addition, predicted polysaccharide biosynthesis gene clusters on the genome sequence, and the high capacity for adhesion, invasion, and cytotoxicity to animal cells confirmed it has pathogenicity on animals. In conclusion, we isolated and identified a novel phytopathogen Erwinia sorbitola sp. nov. in ruddy shelducks. A predefined pathogen is beneficial for preventing from suffering potential economic losses caused by this new pathogen.
Subject(s)
Ducks , Erwinia , Animals , Humans , Phylogeny , Feces , AnxietyABSTRACT
Emerging zoonoses of wildlife origin caused by previously unknown agents are one of the most important challenges for human health. The Qinghai-Tibet Plateau represents a unique ecological niche with diverse wildlife that harbours several human pathogens and numerous previously uncharacterized pathogens. In this study, we identified and characterized a novel arenavirus (namely, plateau pika virus, PPV) from plateau pikas (Ochotona curzoniae) on the Qinghai-Tibet Plateau by virome analysis. Isolated PPV strains could replicate in several mammalian cells. We further investigated PPV pathogenesis using animal models. PPV administered via an intraventricular route caused trembling and sudden death in IFNαßR-/- mice, and pathological inflammatory lesions in brain tissue were observed. According to a retrospective serological survey in the geographical region where PPV was isolated, PPV-specific IgG antibodies were detected in 8 (2.4%) of 335 outpatients with available sera. Phylogenetic analyses revealed that this virus was clearly separated from previously reported New and Old World mammarenaviruses. Under the co-speciation framework, the estimated divergence time of PPV was 77-88 million years ago (MYA), earlier than that of OW and NW mammarenaviruses (26-34 MYA).
Subject(s)
Arenaviridae , Lagomorpha , Animals , Humans , Mice , Arenaviridae/genetics , Phylogeny , Retrospective Studies , Tibet , Animals, WildABSTRACT
The vital functions of extracellular polymeric substances (EPS) have been well recognized in bioleaching of sulfide ores. However, no report is available about the role of EPS in bioleaching of spent catalyst. To completely and deeply understand the functions of EPS in bioleaching of spent catalyst, the generation behavior of EPS at various pulp densities during bioleaching was characterized by three-dimensional excitation-emission matrix (3DEEM), and its relevance with bioleaching performance and process parameters were analyzed using mathematical means. The results showed that the EPS contain humus-like substances as main component (>70%) and protein-like substances as minor component (<30%). Both total EPS and humus-like substances mainly keep growing over the whole duration of bioleaching at low pulp density of 5.0% or lower; whereas total EPS and humus-like fraction keep declining at high pulp density of 7.5% or higher. Among the total EPS and its components, humus-like substances only have a positive significant correlation with bioleaching efficiencies of both Co and Mo and affect bioleaching process more greatly due to greater correlation coefficient. Biofilm appears at the spent catalyst surface under 2.5% of pulp density mediated by EPS while no biofilm occurs at 10% of pulp density due to shortage of EPS, accounting for the great difference in bioleaching efficiencies between high and low pulp densities which are 48.3% for Mo and 50.0% for Co at 10% of pulp density as well as 75.9% for Mo and 78.8% for Co at 2.5% of pulp density, respectively.
Subject(s)
Extracellular Polymeric Substance Matrix , Petroleum , Biofilms , Catalysis , MetalsABSTRACT
Two unknown Gram-stain-positive, catalase- and oxidasenegative, non-motile, and coccus-shaped bacteria, designated MN-17T and MN-09, were isolated from yaks faeces (Bos grunniens) in the Qinghai-Tibet Plateau of China. 16S rRNA gene sequence-based comparative analyses revealed that the two strains were grouped within the genus Vagococcus, displaying the highest similarity with Vagococcus xieshaowenii CGMCC 1.16436T (98.6%) and Vagococcus elongatus CCUG 51432T (96.4%). Both strains grew optimally at 37°C and pH 7.0 in the presence of 0.5% (w/v) NaCl. The complete genome of MN-17T comprises 2,085 putative genes with a total of 2,190,262 bp and an average G + C content of 36.7 mol%. The major fatty acids were C16:0 (31.2%), C14:0 (28.5%), and C18:1ω9c (13.0%); the predominant respiratory quinone was MK-7 (68.8%); the peptidoglycan type was A4α(L-Lys-D-Asp); and the major polar lipid was diphosphatidylglycerol. Together, these supported the affiliation of strain MN-17T to the genus Vagococcus. In silico DNA-DNA hybridization and the average nucleotide identity values between MN-17T and all recognized species in the genus were 21.6-26.1% and 70.7-83.0%, respectively. MN-17T produced acid from D-cellobiose, D-fructose, glycerol, D-glucose, N-acetyl-glucosamine, gentiobiose, D-mannose, D-maltose, D-ribose, D-saccharose, salicin, D-trehalose, and D-xylose. These results distinguished MN-17T and MN-09 from closely related species in Vagococcus. Thus, we propose that strains MN-17T and MN-09 represent a novel species in the genus Vagococcus, with the name Vagococcus zengguangii sp. The type strain is MN-17T (= CGMCC 1.16726T = GDMCC 1.1589T = JCM 33478T).
Subject(s)
Enterococcaceae/isolation & purification , Feces/microbiology , Animals , Bacterial Typing Techniques , Base Composition , Cattle , DNA, Bacterial/genetics , Enterococcaceae/classification , Enterococcaceae/genetics , Enterococcaceae/metabolism , Fatty Acids/metabolism , Genome, Bacterial , RNA, Ribosomal, 16S/geneticsABSTRACT
To investigate the function of CTCF and understand the pathogenesis of tumors better, we produced rabbit polyclonal antibody of human transcription factor CTCF protein and detected its expression in several kinds of human cancer cells and tissues. GST fusion protein of human CTCF-N domain was purified by GSTrap-FF affinity chromatography and was successfully expressed under induction of IPTG in E. coli BL21. Western blotting analysis demonstrated that the polyclonal antibody can recognize the endogenous CTCF from HepG2, MCF-7 and HeLa cells specifically. The produced antibodies can be used for gene expression regulation and tissue distribution study at protein level.
Subject(s)
Antibodies/metabolism , Recombinant Fusion Proteins/immunology , Repressor Proteins/biosynthesis , Repressor Proteins/immunology , Animals , Antibodies/immunology , CCCTC-Binding Factor , Escherichia coli/genetics , Escherichia coli/metabolism , Genetic Vectors/genetics , Glutathione Transferase/biosynthesis , Glutathione Transferase/genetics , HeLa Cells , Hep G2 Cells , Humans , Rabbits , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/genetics , Repressor Proteins/geneticsABSTRACT
A facile one-pot process for synergistic conversion of coal char and methane is conducted by employing K2CO3 as the catalyst. Besides syngas production, valuable carbon products are obtained and used to serve for supercapacitor electrodes. Effect of the operating parameters (including the catalyst dosage, gas feed flow rate, reaction temperature and time) is evaluated on electrochemical performance of the as-prepared carbon. The appropriate surface and structural properties enable the prepared carbon electrode to have a remarkable capacitive performance, along with a specific capacitance up to 125 F/g at a scan rate of 5 mV/s and 133 F/g at a current density of 1 A/g. According to the potential capacitive contribution of the carbon species, the high capacitive performance is mainly attributed to formation and growth of abundant carbon fibers in the one-pot process.
ABSTRACT
OBJECTIVE: To explore possible relationship among expression of human high density lipoprotein binding protein(VIGILIN), H19 and the insulin-like growth factor 2 (IGF2) mRNA in HepG2 cell cycle and investigate the role of VIGILIN in controlling imprinting genes of H19 and IGF2 mRNA expression. METHODS: We investigated time course cell cycle distribution of HepG2 cells by FACS, analyzed VIGILIN, H19 and IGF2 mRNA expression at the indicated times using RT-PCR, RNAi and real-time PCR. RESULTS: Cell-cycle of HepG2 cells was approximately 20 h. 0 h-9 h and 20 h-28 h, 9 h-20 h and 28 h-39 h were S-phase and G2/M-G1-phase, respectively. Firstly, cells were synchronized by serum-starvation for 24 h. As expected, VIGILIN transcription was up-regulated with expression peaks at 20 h and 60 h after serum stimulating by the addition of 10% fetal calf serum. In parallel, H19 mRNA had a high expression level at 6 h and 43 h, and IGF2 mRNA was also increasing with cell-cycle. The expression profiles of human VIGILIN, H19, and IGF2 mRNA were ascending with cell-cycle. In addition, the knock-down of VIGILIN expression by transfecting HepG2 cells with shRNA expression plasmid pSIREN-VIG inhibited the expression of human VIGILIN, which led to the expression of H19 mRNA decrease by 12.08%, and IGF2 mRNA increase by 30.13%. CONCLUSION: The expression of VIGILIN and H19 mRNA was the cell-cycle dependent and had something to do with each other. The results clearly shed light on the roles of VIGILIN in controlling expression of the imprinted H19 and IGF2 genes.
Subject(s)
Genomic Imprinting , Insulin-Like Growth Factor II/genetics , Liver Neoplasms/genetics , RNA, Untranslated/genetics , RNA-Binding Proteins/genetics , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Cell Cycle , Gene Expression Regulation, Neoplastic , Hep G2 Cells , Humans , Insulin-Like Growth Factor II/metabolism , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , RNA Interference , RNA, Long Noncoding , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/genetics , RNA, Untranslated/metabolism , RNA-Binding Proteins/metabolismABSTRACT
OBJECTIVE: To investigate the demethylation effect of CDP on P16 and E-CADHERIN genes. METHODS: Breast cancer cell lines T47D and MDA-MB-435 were treated with CDP and DNA methyltransferase inhibitor 5-azacytidine (5-aza-C). The methylation of P16 and E-CADHERIN gene promoters were measured by methylation-specific PCR (MSP). The RNA transcription was determined by reverse transcription-PCR(RT-PCR). RESULTS: 1) The methylation-specific fragments of P16 gene promoter existed in T47D cells after 25, 50 and 75 micromol/L of CDP treatment for 6 days. An absolute demethylation on P16 gene occurred after treatment with 100 micromol/L of CDP. The unmethylation-specific fragments appeared in T47D cells after being treated with 25, 50, 75 and 100 micromol/L of CDP for 6 days. The RNA expression of P16 was detected after treatment with 75 and 100 micromol/L of CDP. 2) After being treated with 50 micromol/L of CDP, the methylation-specific fragments of CpG island in P16 gene promoter still existed in T47D cells. The unmethylation-specific fragments in T47D cells started to appear after 24 hours of treatment and lasted until 144 hour of treatment. The RNA expression was detected after 144 hours of treatment. 3) The demethylation on E-CADHERIN gene and genomic DNA or RNA transcription were not detected in MDA-MB-435 cells. CONCLUSION: CDP has concentration- and time-dependent demethylation effect on P16 gene in T47D cells, but not on E-CADHERIN gene in MDA-MB-435 cells, which indicates that CDP has substantial diversity in molecular activities.
Subject(s)
Breast Neoplasms/genetics , Cadherins/genetics , DNA Methylation/drug effects , Drugs, Chinese Herbal/pharmacology , Genes, p16 , Azacitidine/pharmacology , Breast Neoplasms/pathology , Cell Line, Tumor , CpG Islands/genetics , HumansABSTRACT
OBJECTIVE: To construct a shRNA eucharyotic expression plasmid against human VIGILIN and explore possible relation between human VIGILIN and HepG2 cell cycle. METHODS: We constructed the shRNA eucharyotic expression plasmid targeted human VIGILIN, and transfected HepG2 cells with shRNA expression plasmid pSIREN-VIG, then determined the expression of VIGILIN mRNA and protein in HepG2 cells by RT-PCR and Western-blot, analysed alteration of cell cycle using FACS. RESULTS: The plasmid pSIREN-VIG can effectively and specifically inhibit the expression of human VIGILIN. After transfection 48 hours, the expression of VIGILIN was significantly decreased. Due to knockdown of human VIGILIN, cell cycle is impaired and cells are arrested in G2/M phase. The proportion of G2/M phase of all groups were listed as: C group (untreated wild HepG2 cells) 2.4%, M group (HepG2 cells treated with transfection reagent) 4.9%, G group (HepG2 cells transfected with pSIREN-GFP) 6.5% and V group (HepG2 cells transfected with pSIREN-VIG) 9.4%. CONCLUSION: We have successfully constructed a shRNA expression plasmid which could effectively and specifically inhibit the expression of human VIGILIN.
Subject(s)
Cell Cycle/genetics , RNA Interference , RNA, Small Interfering/genetics , RNA-Binding Proteins/genetics , Base Sequence , Blotting, Western , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Cell Cycle/physiology , Cell Line, Tumor , Flow Cytometry , Humans , Liver Neoplasms/genetics , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Molecular Sequence Data , Plasmids/genetics , RNA-Binding Proteins/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
CCCTC-binding factor (CTCF), a highly conserved zinc finger protein, is a master organizer of genome spatial organization and has multiple functions in gene regulation. Mounting evidence indicates that CTCF regulates the imprinted genes Igf2 and H19 by organizing chromatin at the Igf2/H19 locus, although the mechanism by which CTCF carries out this function is not fully understood. By yeast two-hybrid screening, we identified vigilin, a multi-KH-domain protein, as a new partner of CTCF. Subsequent coimmunoprecipitation and glutathione S-transferase pulldown experiments confirmed that vigilin interacts with CTCF. Moreover, vigilin is present at several known CTCF target sites, such as the promoter regions of c-myc and BRCA1, the locus control region of ß-globin, and several regions within the Igf2/H19 locus. In vivo depletion of vigilin did not affect CTCF binding; however, knockdown of CTCF reduced vigilin binding to the H19 imprinting control region. Furthermore, ectopic expression of vigilin significantly downregulated Igf2 and upregulated H19, whereas depletion of vigilin upregulated Igf2 and downregulated H19, in HepG2, CNE1 and HeLa cells. These results reveal the functional relevance of vigilin and CTCF, and show that the CTCF-vigilin complex contributes to regulation of Igf2/H19.