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1.
Abolghasem Abbasi Kejani Sayed Ali Hosseini Tafreshi Sayed Mojtaba Khayyam Nekouei Mohammad Reza Mofid 《Molecular biology reports》2010,37(2):797-800
Improved and efficient methods were developed for isolating high quality DNA and RNA from different sources of Iranian Yew
(Taxus baccata L.). The methods were based on CTAB extraction buffer added with high levels of polyvinylpyrrolidone (PVP) and β-mercaptoethanol to properly remove polysaccharides and prevent oxidation of phenolics. The pellets obtained by ethanol precipitation
were washed only with Chloroform: isoamyl alcohol (24:1). So, we could successfully eliminate the dangerous phenol/chloroform
extraction steps from the isolation procedure. Both spectrophotometric (A260/A280 and A260/A230 ratios) and agarose electrophoresis analysis of isolated nucleic acids (DNA and RNA) indicated good results. DNA with the
average yield of 100–300 μg/g leaf and stem tissue and total RNA with an average yield of 20–30 μg/g cell culture and 80–100 μg/g
leaf and stem tissue of Iranian yew could be obtained. Successful amplification of pam and pds by PCR and RT-PCR, showed the integrity of isolated DNA and RNA, respectively. 相似文献
2.
Although resistance of microorganisms to Hg(II) salts has been widely investigated and resistant strains have been reported
from many eubacterial genera, there are few reports of mercuric ion resistance in extremophilic microorganisms. Moderately
thermophilic mercury resistant bacteria were selected by growth at 62 °C on Luria agar containing HgCl2. Sequence analysis of 16S rRNA genes of two isolates showed the closest matches to be with Bacillus pallidus and Ureibacillus thermosphaericus. Minimum inhibitory concentration (MIC) values for HgCl2 were 80 μg/ml and 30 μg/ml for these isolates, respectively, compared to 10 μg/ml for B. pallidus H12 DSM3670, a mercury-sensitive control. The best-characterised mercury-resistant Bacillus strain, B. cereus RC607, had an MIC of 60 μg/ml. The new isolates had negligible mercuric reductase activity but removed Hg from the medium
by the formation of a black precipitate, identified as HgS by X-ray powder diffraction analysis. No volatile H2S was detected in the headspace of cultures in the absence or presence of Hg2+, and it is suggested that a new mechanism of Hg tolerance, based on the production of non-volatile thiol species, may have
potential for decontamination of solutions containing Hg2+ without production of toxic volatile H2S. 相似文献
3.
Changhai Wang Yiyun Wang Qiao Su Xiaorong Gao 《Biotechnology and Bioprocess Engineering》2007,12(2):180-183
A transient expression system for a unicellular marine green alga,Chlorella sp.MACC/C95, was developed using a reporter GUS gene coded for by plasmid pBI121. The results demonstrated a high transformation efficiency
could be achieved by using electroporation to deliver DNA into intact cells and the CaMV35S promoter to drive the foreign
gene expression inChlorella sp.MACC/C95. The use of a carrier DNA coupled with osmosis treatment improved the transformation efficiency, while linearization of the
plasmid had minor effects. Investigation of the effects of DNA concentration and growth phases ofChlorella sp.MACC/C95 on transformation efficiency indicated that the highest level of transient expression was observed when 6 μg mL−1 of plasmid DNA and cells 2–6 days old were used. 相似文献
4.
DNA extraction is difficult in many plants because of metabolites that interfere with DNA isolation procedures and subsequent
applications such as DNA restriction, amplification, and cloning. We developed the first reliable and efficient method for
isolatingVictoria amazonica genomic DNA that is free from polysaccharides and polyphenols. This protocol uses 1.5 M NaCl, 2% polyvinylpyrrolidone (PVP)
(Mr 1000), 5% mercaptoethanol, 0.12% sodium sulfite, and an incubation at 65°C for 4 h. The purity of isolated genomic DNA
was confirmed by means of high-performance liquid chromatography (HPLC) profile and spectrophotometric analyses (A260/230 ratio of 1.836, A260/280 of 1.842). DNA was obtained in the amount of 387 μg per gram of leaf material, and it proved amenable to restriction digestion. 相似文献
5.
Alessandra de Santana Braga Barbosa Ribeiro Cláudio Carlos da Silva Flávia de Castro Pereira Aliny Pereira de Lima Cesar Augusto Sam Tiago Vilanova-Costa Simone Santos Aguiar Luiz Alfredo Pavanin Aparecido Divino da Cruz Elisângela de Paula Silveira-Lacerda 《Biological trace element research》2009,130(3):249-261
Chemotherapeutic agents play an important role in cancer treatment mostly due their systemic action on human organism allowing
access to liquid tumors and even metastases. Among these drugs, ruthenium compounds have been showing promising results to
treat tumors and represent an important development of new antitumor therapy. This study presents the evaluation of cis-(dichloro)tetraammineruthenium(III) chloride, cis-[RuCl2(NH3)4]Cl, genotoxic effects using human peripheral blood lymphocytes cultured in vitro. Mitotic index (MI), chromosome aberrations
(CA), and DNA damage using the comet assay were analyzed. MI in human peripheral blood lymphocyte cultures treated with 1,
10, 100, and 1,000 μg mL−1
cis-[RuCl2(NH3)4]Cl were 5.9%, 4.6%, 3.9%, and 0%, respectively. Doxorubicin chloridate was used as the positive control. CA derived from
1, 10, and 100 μg mL−1 concentrations were defined as spontaneous when compared with the negative control, and at the concentration of 1,000 μg
mL−1, the cell cycle was inhibited (IM = 0%). Results obtained for the comet assay using cis-[RuCl2(NH3)4]Cl suggest that this compound has no genotoxic activity against cultured human peripheral blood lymphocytes. 相似文献
6.
Hongzhen Zhang Fengli Zhang Zhiyong Li 《World journal of microbiology & biotechnology》2009,25(7):1267-1274
Gene cloning, optimized production and property of marine lipase from Bacillus pumilus B106 associated with South China Sea sponge Halichondria rugosa were investigated in this paper. A lipase gene with whole ORF encoding 215 amino acids was obtained by PCR, protein domain
prediction suggested that the deduced lipase belongs to α/β hydrolases family. Based on single factor Seriatim-Factorial test
and Plackett–Burman experimental design, the optimal medium consisted of (per l) 12.5 ml maize oil, 5.0 g beef extract, 2.0 g
PO4
3− (0.6 g KH2PO4, 1.4 g K2HPO4), 17.15 g Mg2+, 5.0 g yeast extract, 2.282 g CaCl2 and 5.0 ml Tween80 with artificial sea water. Using this optimum medium, lipase activity and cell concentration were increased
by 3.54- and 1.31-fold over that of the basal medium, respectively. This lipase showed tolerance to high salinity, pH and
temperature. About 10–20% methanol exhibited a stimulatory effect on the lipase activity, while activity was inhibited by
30–40% methanol, 2-propanol, DMSO, and ethanol. This study provides a valuable resource for marine lipase production and extends
our understanding of the possible role of sponge-associated bacteria in the biotransformation of chemical compounds for the
sponge host. 相似文献
7.
The biokinetics of glucose metabolism were evaluated in Aeromonas hydrophila during growth in an anaerobic biosystem. After approx 34 h growth, A. hydrophila metabolized 5,000 mg glucose l−1 into the end-products ethanol, acetate, succinate and formate. The maximum growth rate, μ
m, half saturation coefficients, K
s, microbial yield coefficient, Y, cell mass decay rate coefficient, k
d, and substrate inhibition coefficient, K
si were 0.25 ± 0.03 h−1, 118 ± 31 mg glucose l−1, 0.12 μg DNA mg glucose−1, 0.01 h−1, and 3,108 ± 1,152 mg glucose l−1, respectively. These data were used to predict the performance of a continuous growth system with an influent glucose concentration
of 5,000 mg l−1. Results of the analysis suggest that A. hydrophila will metabolize glucose at greater than 95% efficiency when hydraulic retention times (HRTs) exceed 7 h, whereas the culture
is at risk of washing out at an HRT of 6.7 h. 相似文献
8.
Yemendzhiev H Gerginova M Krastanov A Stoilova I Alexieva Z 《Journal of industrial microbiology & biotechnology》2008,35(11):1309-1312
Trametes versicolor 1 was shown to grow on phenol as its sole carbon and energy source. The culture growth and degradation ability dependence
on culture medium pH value was observed. The optimal pH value of a liquid Czapek salt medium was 6.5. The investigated strain
utilized completely 0.5 g/l phenol in 6 days. The dynamics of the phenol degradation process was investigated. The process
was characterized by specific growth rate μmax 0.33 h−1, metabolic coefficient k = 4.4, yield coefficient Y
x/s
= 0.23 and rate of degradation Q = 0.506 h−1. The intracellular activities of phenol hydroxylase (0.333 U/mg protein) and cis,cis-muconate lactonizing enzyme (0.41 U/mg protein) were demonstrated for the first time in this fungus. In an attempt to estimate
the occurrence of gene sequences in T. versicolor 1 related to phenol degradation pathway a dot blot analysis with total DNA isolated from this strain was performed. Two synthetic
oligonucleotides were used as hybridizing probes. One of the probes was homologous to the 5′end of phyA gene coding for phenol hydroxylase in Trichosporon cutaneum ATCC 46490. The other probe was created on the basis of cis,cis-muconate lactonizing enzyme coding gene in T. cutaneum ATCC 58094. The results of these investigations showed that T. versicolor 1 may carry genes similar to those of Trichosporon cutaneum capable to degrade phenol. 相似文献
9.
Ki Jun Jeong Hyun Sook Lee Sang Yup Lee Yong Keun Chang 《Biotechnology and Bioprocess Engineering》1998,3(1):48-49
A protocol for the transformation ofKlebsiella oxytoca by electroporation was developed. Preparation of competent cells at early exponential phase was most critical to obtain a
high transformation efficiency. The highest efficiency of 1.6 × 106 transformants per μg DNA (pBR 322) could be obtained by electroporation ofK. oxytoca cells prepared at the OD600 of 0.2 with 1.25 μg DNA at the filed strength of 2.5 kV, the parallel resistance of 200 Ω and capacitance of 25 μF. 相似文献
10.
Ruirong Yi Takashi Tachikawa Hiroyuki Mukaiyama Yusuke Mochida Mariko Ishikawa Tadanori Aimi 《Mycoscience》2009,50(2):123-129
We cloned a gene encoding the succinate dehydrogenase iron-sulfur protein subunit (sip) from a bipolar mushroom, Pholiota microspora, and introduced a point mutation that confers carboxin resistance into this gene. Using this homologous selective marker
and also a heterologous drug selective marker, the hygromycin B phosphotransferase gene (hph), we successfully constructed a DNA-mediated transformation system in P. microspora. Both these selection markers have high transformation efficiency: the efficiency of carboxin resistance transformation was
about 88.8 transformants/μg pMBsip2 DNA using 5 × 106 protoplasts in regeneration plates containing 1.0 μg/ml carboxin, and the efficiency of hygromycin B resistance transformation
was about 122.4 transformants/μg pMBhph1 DNA using 5 × 106 protoplasts in regeneration plates containing 150 μg/ml hygromycin B. Southern hybridization analysis showed that the introduced
sequence (mutant sip or hph) was integrated into the chromosomal DNA in these transformants with a copy number of one or more. 相似文献
11.
Fusarium toxins are secondary metabolites produced byfungi of these genera in many commodities under certain conditions. A study was carried out to investigate the co-occurrence of
zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FB1 and FB2) in 52 samples of mixed-feed for poultry contaminated withFusarium verticillioides. The zearalenone and deoxynivalenol were checked using immunoaffinity column and the extraction of fumonisin was performed
by strong anion exchange (SAX) solid phase column. Detection and quantification were determined by high performance liquid
chromatography (HPLC). The limit of detection was 5 μg/kg for ZEN, 100 μg/kg for DON and 50 and 100 μg/kg for FB1 and FB2 respectively.Fusarium toxins were detected in 20 samples. Sixteen samples were positive for ZEN (30.7%) presenting levels that ranged from 7.4
μg/kg to 61.4 μg/kg (mean=27.0 μg/kg). 13.5% of the samples presented contaminations of DON, with levels ranging from 100.0
μg/kg to 253 μg/kg (mean=l18.07 μg/kg). FB1 was detected in 19.2% of samples, with levels ranging from 50.0 μg/kg to 110.0 μg/kg (mean=73.6 μg/kg). FB2 was not detected
in any sample. In positive samples simultaneously contamination with two or three mycotoxins were detected in 9 of them (17.3%). 相似文献
12.
Damayanti De Ananda Mukhopadhyay Ranadhir Chakraborty 《World journal of microbiology & biotechnology》2008,24(11):2727-2729
Enterobacter sp. was isolated from the diseased and dead caterpillars of the tea leaf roller (Caloptilia theivora) from the Darjeeling foothill region. When the vegetative form of the bacterium was applied via food, mortality of C. theivora showed an LC50 value at 363.1 μg/ml (bacterial wt./vol. of water) with fiducial limits 363.25 and 362.94 μg/ml respectively. The LT50 values for C. theivora were 6 days for 100 μg/ml, 5.96 days for 300 μg/ml, 5.81 days for 500 μg/ml, 4.96 days for 750 μg/ml and 4.61 days for 1,000 μg/ml
concentrations. The finding would enable one to contemplate development of a microbial pesticide using this novel Enterobacter sp. DD01 for control of the leaf rolling pest. 相似文献
13.
Lakkana Laopaiboon Pornthap Thanonkeo Prasit Jaisil Pattana Laopaiboon 《World journal of microbiology & biotechnology》2007,23(10):1497-1501
Sweet sorghum juice supplemented with 0.5% ammonium sulphate was used as a substrate for ethanol production by Saccharomyces cerevisiae TISTR 5048. In batch fermentation, kinetic parameters for ethanol production depended on initial cell and sugar concentrations.
The optimum initial cell and sugar concentrations in the batch fermentation were 1 × 108 cells ml−1 and 24 °Bx respectively. At these conditions, ethanol concentration produced (P), yield (Y
ps) and productivity (Q
p
) were 100 g l−1, 0.42 g g−1 and 1.67 g l−1 h−1 respectively. In fed-batch fermentation, the optimum substrate feeding strategy for ethanol production at the initial sugar
concentration of 24 °Bx was one-time substrate feeding, where P, Y
ps and Q
p
were 120 g l−1, 0.48 g g−1 and 1.11 g l−1 h−1 respectively. These findings suggest that fed-batch fermentation improves the efficiency of ethanol production in terms of
ethanol concentration and product yield. 相似文献
14.
F. W. Wang Z. M. Hou C. R. Wang P. Li D. H. Shi 《World journal of microbiology & biotechnology》2008,24(10):2143-2147
The metabolites of endophytic fungus Penicillium sp. from the leaf of Hopea hainanensis were reported for the first time. By bioassay-guided fractionation, the EtOAc extract of a solid-matrix steady culture of
this fungus afforded six compounds, which were identified through a combination of spectral and chemical methods (IR, MS,
1H- and 13C-NMR) to be monomethylsulochrin (1), rhizoctonic acid (2), asperfumoid (3), physcion (4), 7,8-dimethyl-iso-alloxazine (5) and 3,5-dichloro-p-anisic acid (6). Compounds 2, 3 and 6 were obtained from Penicillium sp. for the first time. All of the six isolates were subjected to in vitro bioactive assays including antifungal action against
three human pathogenic fungi Candida albicans, Trichophyton rubrum and Aspergillus niger and cytotoxic activity against the human nasopharyngeal epidermoid tumor KB cell line and human liver cancer HepG2 cell line.
As a result, compounds 2–4 and 6 inhibited the growth of C. albicans with MICs of 40.0, 20.0, 50.0 and 15.0 μg/ml, respectively and the compound 6 showed growth inhibition against A. niger with MICs of 40.0 μg/ml. In addition, compounds 1–3 and 6 exhibited cytotoxic activity against KB cell line with IC50 value of 30.0, 20.0, 20.0, 5.0 μg/ml, respectively and against HepG2 cell line with IC50 value of 30.0, 25.0, 15.0, 10.0 μg/ml, respectively. 相似文献
15.
Leandro Finkler Christine Lamenha Luna-Finkler José Carlos Pinto Tito Livio Moitinho Alves 《World journal of microbiology & biotechnology》2007,23(12):1789-1795
Separation and cells concentration constitute important stages in most biotechnological processes. Particularly, use of flocculation/sedimentation
can improve significantly the extraction of biopolymers accumulated by microorganisms and the biodegradation of xenobiotic
compounds by cell sludge. In this work the use of tannin and aluminum sulphate (Al2(SO4)3) as flocculating agents for concentration of cells of Cupriavidus necator DSM 545 is evaluated. Cells were grown in broth nutrient medium in Erlenmeyer flasks, submitted to orbital agitation of 160 rpm
at 30 °C for 21 h. The optimal concentrations of flocculating agents, as determined with a standard jar test method, were
equal to 2,800 mg/L for tannin and 800 mg/L for Al2(SO4)3, allowing for recovery of 95% of the cells in both cases. Obtained flocs presented density and average diameter of 1.03 g/mL ± 0.01 g/mL
and 158 μm ± 19 μm for tannin and of 1.05 g/mL ± 0.01 g/mL and 146 μm ± 14 μm for Al2(SO4)3, respectively. Batch settling tests were performed in order to determine the operational capacity of continuous settlers
to be used for separation of the investigated flocculent suspensions. Finally, cultivation of cells using flocs as inoculum
indicated that the cells remained viable after flocculation with usage of the optimum flocculating agent concentrations. 相似文献
16.
Four temperature treatments were studied in the climate controlled growth chambers of the Georgia Envirotron: 25/20, 30/25,
35/30, and 40/35 °C during 14/10 h light/dark cycle. For the first growth stage (V3-5), the highest net photosynthetic rate
(P
N) of sweet corn was found for the lowest temperature of 28–34 μmol m−2 s−1 while the P
N for the highest temperature treatment was 50–60 % lower. We detected a gradual decline of about 1 P
N unit per 1 °C increase in temperature. Maximum transpiration rate (E) fluctuated between 0.36 and 0.54 mm h−1 (≈5.0–6.5 mm d−1) for the high temperature treatment and the minimum E fluctuated between 0.25 and 0.36 mm h−1 (≈3.5–5.0 mm d−1) for the low temperature treatment. Cumulative CO2 fixation of the 40/35 °C treatment was 33.7 g m−2 d−1 and it increased by about 50 % as temperature declined. The corresponding water use efficiency (WUE) decreased from 14 to
5 g(CO2) kg−1(H2O) for the lowest and highest temperature treatments, respectively. Three main factors affected WUE, P
N, and E of Zea: the high temperature which reduced P
N, vapor pressure deficit (VPD) that was directly related to E but did not affect P
N, and quasi stem conductance (QC) that was directly related to P
N but did not affect E. As a result, WUE of the 25/20 °C temperature treatment was almost three times larger than that of 40/35 °C temperature treatment. 相似文献
17.
Domínguez-Bocanegra AR Ponce-Noyola T Torres-Muñoz JA 《Applied microbiology and biotechnology》2007,75(4):783-791
Phaffia rhodozyma (now Xanthophyllomyces dendrorhous) and Haematococcus pluvialis are known as the major prominent microorganisms able to synthesize astaxanthin natural pigment. Important research efforts
have been made to determine optimal conditions for astaxanthin synthesis. When the focus is on astaxanthin production, the
maximal reported value of 9.2 mg/g cell is obtained within H. pluvialis grown on BAR medium, under continuous illumination (345 μmol photon m−2 s−1) and without aeration. Whereas fermentation by mutated R1 yeast grown on coconut milk produced 1,850 μg/g yeast. However,
when looking at astaxanthin productivity, the picture is slightly different. The figures obtained with P. rhodozyma are rather similar to those of H. pluvialis. Maximal reported values are 170 μg/g yeast per day with a wild yeast strain and 370 μg/g yeast per day with mutated R1 yeast.
In the case of H. pluvialis, maximal values ranged from 290 to 428 μg/g cell per day depending on the media (BG-11 or BAR), light intensity (177 μmol
photon m−2 s−1), aeration, etc. The main aim of this work was to examine how astaxanthin synthesis, by P. rhodozyma and H. pluvialis, could be compared. The study is based on previous works by the authors where pigment productions have been reported. 相似文献
18.
Huang HY Chen YG Wang YX Liu JH Tang SK Peng Q Wen ML Yu H Cui XL 《Extremophiles : life under extreme conditions》2008,12(6):829-835
A novel Gram-negative, slightly halophilic, catalase- and oxidase-positive, obligately aerobic bacterium, strain YIM-C248T, was isolated from a sediment sample collected from a salt-lake in the Qaidam Basin in Qinghai, north-west China. Cells were
non-sporulating short rods, occurring singly or as doublets, motile with peritrichous flagella. Growth occurred with 1–15%
(w/v) NaCl [optimum 2–4% (w/v) NaCl], at pH 6.0–10.0 (optimum pH 7.5) and at 4–35°C (optimum 25–30°C). The major cellular
fatty acids were C18:1
ω7c, C12:0 3-OH, cyclo C19:0
ω8c, C16:0 and C16:1. The predominant respiratory quinone was Q-9 and the genomic DNA G + C content was 58.6 mol%. Phylogenetic analysis based
on 16S rRNA gene sequences indicated that strain YIM-C248T should be assigned to the genus Halomonas. The sequence similarities between the isolate and the type strains of members of the genus Halomonas were in the range of 92.5–97.5%. The combination of phylogenetic analysis, DNA–DNA hybridization data, phenotypic characteristics
and chemotaxonomic differences supported the view that strain YIM-C248T represents a new species of the genus Halomonas, for which the name Halomonas sediminis sp. nov. is proposed, with YIM-C248T (=CCTCC AA 207031 = KCTC 22167) as the type strain.
The GenBank/EMBL/DBBJ accession number for the 16S rRNA gene sequence of strain YIM-C248T is EU135707. 相似文献
19.
The endochitinase DNA and cDNA from Trichoderma sp. were cloned, sequenced and expressed. The cloned DNA and cDNA sequences were 1,476 and 1,275 bp in length, respectively.
There were three introns in DNA sequence in comparison with the cDNA sequence. The endochitinase protein contained three regions:
the signal peptide, the prepro-region and the mature protein region. The gene fragment encoding the mature endochitinase was
ligated into the expression vector pET-28a+, yielding pET-1. The plasmid pET-1 was transformed into the Escherichia coli BL21 (DE3). The clone bearing pET-1 was picked and cultured at 30°C for the expression of endochitinase. SDS-PAGE analysis
showed that the endochitinase was expressed in the periplasmic space and the purified protein showed a single band. The activity
of 70.2 U/mg was obtained from the cellular extract of the recombinant strain. The activity of endochitinase was 2.5-fold
higher at 24 h than at 16 h in the periplasmic space. The optimal pH and temperature of the recombinant endochitinase were
determined to be 7.0 and 35°C, respectively. It was relatively stable within the pH range of 5–8. Significant activity stimulation
by 1 mM Mg2+ and 5 mM Fe2+ and inhibition by 5 mM Co2+ and 5 mM Hg2+ were observed. The kinetic constants Km, Vmax and Kcat for the hydrolysis of the colloidal chitin were 1.5 mM, 1.37 μmol min−1 and 6.23 min−1, respectively. 相似文献
20.
Metabolically-engineered Escherichia coli strains were developed by cloning poly-γ-glutamic acid (γ-PGA) biosynthesis genes, consisting of pgsB, pgsC and pgsA, from Bacillus subtilis The metabolic and regulatory pathways of γ-PGA biosynthesis in E. coli were analyzed by DNA microarray. The inducible trc promoter and a constitutive promoter (PHCE) derived from the d-amino acid aminotransferase (D-AAT) gene of Geobacillus toebii were employed. The constitutive HCE promoter was more efficient than inducible trc promoter for the expression of γ-PGA biosynthesis genes. DNA microarray analysis showed that the expression levels of several NtrC family genes, glnA, glnK, glnG, yhdX, yhdY, yhdZ, amtB, nac, argT and cbl were up-regulated and sucA, B, C, D genes were down-regulated. When (NH4)2SO4 was added at 40 g/l into the feeding solution, the final γ-PGA concentration reached 3.7 g/l in the fed-batch culture of recombinant E. coli/pCOpgs. 相似文献