Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, P...Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 mug/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.展开更多
Delta-12 oleate desaturase gene (FAD2-1) which converts oleic acid into linoleic acid, is the key enzyme determining the fatty acid composition of cottonseed oil. By employing RT-PCR method, full length cDNA of cott...Delta-12 oleate desaturase gene (FAD2-1) which converts oleic acid into linoleic acid, is the key enzyme determining the fatty acid composition of cottonseed oil. By employing RT-PCR method, full length cDNA of cotton delta-12 oleate desat- urase gene GhFAD2-1 containing an open reading frame of 1 158 bp was cloned for constructing RNAi vector. A 515 bp long specific fragment of this gene was se- lected for constructing ihpRNA vector under the control of a seed-specific promoter NAPIN, named pFGC1008-NAPIN-FAD2-1; meanwhile miRNA gene-silencing vector pCAMBIA1302-amiRNA-FAD2-1 targeting GhFAD2-1 was also constructed.展开更多
Highly acidic crude oil is thermally soaked to investigate how the temperature and time involved affect the removal of organic acid in feedstock. Experimental results indicate that thermal treatment is an effective a...Highly acidic crude oil is thermally soaked to investigate how the temperature and time involved affect the removal of organic acid in feedstock. Experimental results indicate that thermal treatment is an effective approach to decreasing acidity and the acid removal rate reaches 80%. Temperature is one of the main factors that determine the acid removal reaction. When the temperature ranges from 420oC to 440oC, the acid removal rate increases with the rise of the reaction temperature, but the increase slows down gradually. At the reaction temperature below 440oC, the long reaction time favors the acid removal. The cracking and polymerization of hydrocarbon molecules take place so that the properties of the crude oil change at the same time when the highly acidic crude is thermally treated.展开更多
A new kind of solvent for deacidification of distillate oils was introduced in this paper. After successful laboratory study this technology had been applied in commercial scale successfully. Compared to traditional c...A new kind of solvent for deacidification of distillate oils was introduced in this paper. After successful laboratory study this technology had been applied in commercial scale successfully. Compared to traditional caustic wash of distillate oils, this technology has a lot of merits, such as the broad range of distillates to be processed, low caustic consumption, recycle of deacidifying agent, absence of waste caustic discharge, and low equipment revamp expenses, which can have promising perspectives for exploitation and application of this technology.展开更多
The use of H2SO4‐,HCl‐,H3PO4‐,and CH3COOH‐activated montmorillonite(Mt)and WOx/H3PO4‐activated Mt as catalysts for the gas‐phase dehydration of glycerol was investigated.The WOx/H3PO4‐activated Mt catalysts wer...The use of H2SO4‐,HCl‐,H3PO4‐,and CH3COOH‐activated montmorillonite(Mt)and WOx/H3PO4‐activated Mt as catalysts for the gas‐phase dehydration of glycerol was investigated.The WOx/H3PO4‐activated Mt catalysts were prepared by an impregnation method using H3PO4‐activated Mt(Mt‐P)as the support.The catalysts were characterized using powder X‐ray diffraction,Fourier‐transform infrared spectroscopy,N2adsorption‐desorption,diffuse reflectance ultraviolet‐visible spectroscopy,temperature‐programmed desorption of NH3,and thermogravimetric analysis.The acid activation of Mt and WOx loaded on Mt‐P affected the strength and number of acid sites arising from H+exchange,the leaching of octahedral Al3+cations from Mt octahedral sheets,and the types of WOx(2.7≤x≤3)species(i.e.,isolated WO4/WO6‐containing clusters,two‐dimensional[WO6]polytungstates,or three‐dimensional WO3crystals).The strong acid sites were weakened,and the weak and medium acid sites were strengthened when the W loading on Mt‐P was12wt%(12%W/Mt‐P).The12%W/Mt‐P catalyst showed the highest catalytic activity.It gave a glycerol conversion of89.6%and an acrolein selectivity of81.8%at320°C.Coke deposition on the surface of the catalyst led to deactivation.展开更多
A facile approach to the preparation of a novel magnetically separable H_5PMo_(10)V_2O_(40)/Fe_3O_4/g-C_3N_4(PMoV/Fe_3O_4/g-C_3N_4) nanocomposite by chemical impregnation is demonstrated.The prepared nanocomposi...A facile approach to the preparation of a novel magnetically separable H_5PMo_(10)V_2O_(40)/Fe_3O_4/g-C_3N_4(PMoV/Fe_3O_4/g-C_3N_4) nanocomposite by chemical impregnation is demonstrated.The prepared nanocomposite was characterized and its acidity was measured by potentiometric titration.PMoV/Fe_3O_4/g-C_3N_4 showed high catalytic activity in the selective oxidative desulfurization of sulfides to their corresponding sulfoxides or sulfones.The catalytic oxidation of a dibenzothiophene(DBT)-containing model oil and that of real oil were also studied under optimized conditions.In addition,the effects of various nitrogen compounds,as well as the use of one- and two-ring aromatic hydrocarbons as co-solvents,on the catalytic removal of sulfur from DBT were investigated.The catalyst was easily separated and could be recovered from the reaction mixture by using an external magnetic field.Additionally,the remaining reactants could be separated from the products by simple decantation if an appropriate solvent was chosen for the extraction.The advantages of this nanocatalyst are its high catalytic activity and reusability;it can be used at least four times without considerable loss of activity.展开更多
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) plays important roles in various cellular processes. A cytosolic GAPDH encoding gene (gpd) of Gracilaria/Gracilariopsis lemaneiformis was cloned and characterized. ...Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) plays important roles in various cellular processes. A cytosolic GAPDH encoding gene (gpd) of Gracilaria/Gracilariopsis lemaneiformis was cloned and characterized. Deduced amino acid sequence of the enzyme of G. lemaneiformis had high homology with those of seven red algae. The 5'-untranslated regions of the GAPDHs encoding genes of these red algae varied greatly. GAPDHs of these red algae shared the highly conserved glyceraldehyde 3-phosphate dehydrogenase active site ASCTTNCL. However, such active site of Cyanidium caldarium was different from those of the other six algae at the last two residues (CL to LF), thus the spatial structure of its GAPDH active center may be different from those of the other six. Phylogenetic analysis indicated that GAPDH of G. lemaneiformis might have undergone an evolution similar to those of Porphyra yezoensis, Chondrus crispus, and Gracilaria verrucosa. C. caldarium had a closer evolutionary relationship with Cyanidioschyzon merolae than with Cyanidium sp. Virtual Northern blot analysis revealed that gpd of G. lemaneiformis expressed constitutively, which suggested that it might be house-keeping and could be adapted as an inner control in gene expression analysis of G. lemaneiformis.展开更多
Accurate quantification of transcripts using quantitative real-time polymerase chain reaction (qPCR) depends on the identification of reliable reference genes for normalization. This study aimed to identify and vali...Accurate quantification of transcripts using quantitative real-time polymerase chain reaction (qPCR) depends on the identification of reliable reference genes for normalization. This study aimed to identify and validate seven reference genes, including actin-2 (ACT-2), elongation factor 1 alpha (EF-1α), elongation factor 1 beta (EF-1β), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin (UBQ), β-tubulin (β-TUB), and 18 S ribosomal RNA, from Crassostrea angulata, a valuable marine bivalve cultured worldwide. Transcript levels of the candidate reference genes were examined using qPCR analysis and showed differential expression patterns in the mantle, gill, adductor muscle, labial palp, visceral mass, hemolymph and gonad tissues. Quantitative data were analyzed using the geNorm software to assess the expression stability of the candidate reference genes, revealing that β-TUB and UBQ were the most stable genes. The commonly used GAPDH and 18S rRNA showed low stability, making them unsuitable candidates in this system. The expression pattern of the G protein β-subunit gene (Gβ) across tissue types was also examined and normalized to the expression of each or both of UBQ andβ-TUB as internal controls. This revealed consistent trends with all three normalization approaches, thus validating the reliability of UBQ and β-TUB as optimal internal controls. The study provides the first validated reference genes for accurate data normalization in transcript profiling in Crassostrea angulata, which will be indispensable for further fimetional genomics studies in this economically valuable marine bivalve.展开更多
An experiment was performed to determine the dietary phosphorus requirement of the young abalone, Haliotis discus hannai. Five semi-purified diets were formulated to provide a series of graded levels of dietary total ...An experiment was performed to determine the dietary phosphorus requirement of the young abalone, Haliotis discus hannai. Five semi-purified diets were formulated to provide a series of graded levels of dietary total phosphorus (0.23%-1.98) from monobasic potassium phosphate (KH2PO4). The brown alga, Laminaria japonica, was used as a control diet. Similar size abalone were distributed in a single-pass, flow-through system using a completely randomized design with six treatments and three replicates each treatment. The abalone were hand-fed to satiation with appropriate diets in excess, once daily at 17:00. The feeding trial was run for 120-d. Survival rate and soft-body to shell ratio (SB/S) were constantly maintained regardless of dietary treatment. However, the weight gain rate (WGR), daily increment in shell length (DISL), muscle RNA to DNA ratio (RNA/DNA), carcass levels of lipid and protein, soft-body alkaline phosphatase (SBAKP), and phosphorus concentrations of whole body (WB) and soft body (SB) were significantly (ANOVA, P<0.05) affected by the dietary phosphorus level. The dietary phosphorus requirements of the abalone were evaluated from the WGR, DISL, and RNA/DNA ratio respectively, by using second-order polynomial regression analysis. Based on these criteria, about 1.0%-1.2% total dietary phosphorus, i.e. 0.9%-1.1% dietary available phosphorus is recommended for the maximum growth of the abalone.展开更多
The carboxyl terminal of sodium oleate has a stronger polarity than that of oleic acid;this terminal is more likely to be dipole polarized and ionically conductive in a microwave field.Sodium oleate was used as the mo...The carboxyl terminal of sodium oleate has a stronger polarity than that of oleic acid;this terminal is more likely to be dipole polarized and ionically conductive in a microwave field.Sodium oleate was used as the model compound to study the decarboxylation of oleic acid leading to hydrocarbon formation via microwave-assisted pyrolysis technology.The pyrolysis gas,liquid,and solid products were precisely analyzed to deduce the mechanism for decarboxylation of sodium oleate.Microwave energy was able to selectively heat the carboxyl terminal of sodium oleate.During decarboxylation,the double bond in the long hydrocarbon chain formed a p-πconjugated system with the carbanion intermediate.The resulting p-πconjugated system was more stable and beneficial to the pyrolysis reaction(decarboxylation,terminal allylation,isomerization,and aromatization).The physical properties of pyrolysis liquid were generally similar to those of diesel fuel,thereby demonstrating the possible use of microwaves for controlling the decarboxylation of sodium oleate in order to manufacture renewable hydrocarbon fuels.展开更多
The purpose of this study is to increase acrolein yield and capability of coking resistance in the reaction of glycerol dehydration to acrolein by assembling metal phosphate supported on HZSM-5 catalyst. The as-prepar...The purpose of this study is to increase acrolein yield and capability of coking resistance in the reaction of glycerol dehydration to acrolein by assembling metal phosphate supported on HZSM-5 catalyst. The as-prepared catalysts were characterized by XRD, SEM, EDS, BET, NH_3-TPD, CO_2-TPD and Py-IR techniques. It was found that metal phosphate species were incorporated into the porous structure of HZSM-5 zeolites, thus influencing the surface and textural physico-chemical properties of the supporters. The alkaline-treated HZSM-5 catalyst promoted the dispersion of phosphate species on the carriers. Moreover, the amount of strong acidity was tremendously improved by adding the different metal hydrophosphates and the catalysts show high catalytic activity. In this present work, the Sn1/4 H_2PO_4/HZSM-5 catalyst exhibited good performance in the catalytic activity and coking resistant ability, which resulted in a high acrolein yield of 83% initially and acrolein yield of 68% after 30 h. The acidity, especially the ratio of strong to weak acid, plays an important role in promoting acrolein yield and stability simultaneously.展开更多
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), initially identified as a glycolytic enzyme and considered as a housekeeping gene, is widely used as an internal control in experiments on proteins, mRNA, and DNA. H...Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), initially identified as a glycolytic enzyme and considered as a housekeeping gene, is widely used as an internal control in experiments on proteins, mRNA, and DNA. However, emerging evidence indicates that GAPDH is implicated in diverse functions independent of its role in energy metabolism; the expression status of GAPDH is also deregulated in various cancer cells. One of the most common effects of GAPDH is its inconsistent role in the determination of cancer cell fate. Furthermore, studies have described GAPDH as a regulator of cell death; other studies have suggested that GAPDH participates in tumor progression and serves as a new therapeutic target. However, related regulatory mechanisms of its numerous cellular functions and deregulated expression levels remain unclear. GAPDH is tightly regulated at transcriptional and pnsttranscriptional levels, which are involved in the regulation of diverse GAPDH functions. Several cancer-related factors, such as insulin, hypoxia inducible factor-1 (HIF-1), p53, nitric oxide (NO), and acetylated histone, not only modulate GAPDH gene expression but also affect protein functions via common pathways. Moreover, posttranslational modifications (PTMs) occurring in GAPDH in cancer cells result in new activities unrelated to the original glycnlytic function of GAPDH. In this review, recent findings related to GAPDH transcriptional regulation and PTMs are summarized. Mechanisms and pathways involved in GAPDH regulation and its different roles in cancer cells are also described.展开更多
The removal of acid compounds (naphthenates) from acidic oil with ionic liquids was systematically investigated. [BMIM]Br-AlCl3 was used to investigate the effect on deacidification of oil. Experimental results show...The removal of acid compounds (naphthenates) from acidic oil with ionic liquids was systematically investigated. [BMIM]Br-AlCl3 was used to investigate the effect on deacidification of oil. Experimental results showed that at a temperature of 323K with a molar ratio of AlCl3 to [BM1M]Br-AlCl3 of 0.2, and a mass ratio of IL to white oil of 4%, the deacidification rate could reach 75.9%. And a reaction time of 4 h was sufficient to achieve the goal. The study on reproducibility of catalytic performance of [BMIM]Br-AlCl3 showed the possibility of using the ionic liquid in the continuous catalytic reaction.展开更多
The method for refining the straight-run diesel fuel was studied in laboratory scale in order to make the acid number of diesel fraction comply with the standard while removing the naphthenic acids contained in diesel...The method for refining the straight-run diesel fuel was studied in laboratory scale in order to make the acid number of diesel fraction comply with the standard while removing the naphthenic acids contained in diesel without causing environmental pollution. After comparing the effect of refining using three solvents, the isopropyl alcohol-HOA was specified as the best solvent. Meanwhile, the relationship between the acid number of diesel fraction and the amount of solvent used and the relationship between the concentration of solvent and temperature and the stability of diesel in terms of its acid number were also investigated. Experimental results had shown that when the mass fraction of the HOA-IPA solvent was 20% at a dosage of 17 mL of the solvent and a temperature of 30℃, the acid number of the refined diesel fraction was 0.015 mg KOH/g with a good stability of acidity in the diesel fraction.展开更多
Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the...Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde- 3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that a-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.展开更多
The aim of the study was to evaluate the impact of organic selenium supplementation (selenium yeast) vs. inorganic selenium (selenium selenite) on the expression of selected defensin genes in milk somatic cells (...The aim of the study was to evaluate the impact of organic selenium supplementation (selenium yeast) vs. inorganic selenium (selenium selenite) on the expression of selected defensin genes in milk somatic cells (MSC) of dairy cows. Sixteen Polish Holstein-Friesian cows in second parity, with similar milk yield and milk composition were randomly divided into two equal groups. At the beginning of experiment, the animals were in the middle of their lactation curve (150, standard deviation (SD) = 26 d) and this experiment lasted 90 d. The basic diet, for both groups, consisted of corn silage, wilted grass silage and concentrates. The diet of control group was supplemented with commercial mineral and vitamin mixture with inorganic selenium and the for the experimental group daily selenium need was covered by the addition of Se-yeast (6 mg Se/cow/d). Milk samples were collected three times during experiment (before experiment; on day 55; after the end the experiment). Total RNA was isolated from milk somatic cells and the levels of transcripts of bovine β1-defensin (DEFB1), β4-defensin (BNBD4), β5-defensin (BNBDS), β10-defensin (BNBD10) and lingual antimicrobial peptide (LAP) were measured with real time PCR, using glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a reference gene. The expressions of almost all studied genes were influenced by environmental conditions (very low and negative temperature). However, BNBD4 (P 〈 0.05) and LAP (P ≤ 0.001 ) genes were influenced by selenium supplementation but in opposite ways, depending on the form of Se. These findings support that selenium is an important factor affecting the mRNA level in MSC, but the effect of the form of selenium might depend on genes.展开更多
文摘Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 mug/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.
文摘Delta-12 oleate desaturase gene (FAD2-1) which converts oleic acid into linoleic acid, is the key enzyme determining the fatty acid composition of cottonseed oil. By employing RT-PCR method, full length cDNA of cotton delta-12 oleate desat- urase gene GhFAD2-1 containing an open reading frame of 1 158 bp was cloned for constructing RNAi vector. A 515 bp long specific fragment of this gene was se- lected for constructing ihpRNA vector under the control of a seed-specific promoter NAPIN, named pFGC1008-NAPIN-FAD2-1; meanwhile miRNA gene-silencing vector pCAMBIA1302-amiRNA-FAD2-1 targeting GhFAD2-1 was also constructed.
文摘Highly acidic crude oil is thermally soaked to investigate how the temperature and time involved affect the removal of organic acid in feedstock. Experimental results indicate that thermal treatment is an effective approach to decreasing acidity and the acid removal rate reaches 80%. Temperature is one of the main factors that determine the acid removal reaction. When the temperature ranges from 420oC to 440oC, the acid removal rate increases with the rise of the reaction temperature, but the increase slows down gradually. At the reaction temperature below 440oC, the long reaction time favors the acid removal. The cracking and polymerization of hydrocarbon molecules take place so that the properties of the crude oil change at the same time when the highly acidic crude is thermally treated.
文摘A new kind of solvent for deacidification of distillate oils was introduced in this paper. After successful laboratory study this technology had been applied in commercial scale successfully. Compared to traditional caustic wash of distillate oils, this technology has a lot of merits, such as the broad range of distillates to be processed, low caustic consumption, recycle of deacidifying agent, absence of waste caustic discharge, and low equipment revamp expenses, which can have promising perspectives for exploitation and application of this technology.
基金supported by the National Natural Science Foundation of China(21373185,41672033,21506188,21404090)the Open Project Programs of Engineering Research Center of Non-metallic Minerals of Zhejiang Province(ZD2015k07)+2 种基金of State Key Laboratory Breeding Base of Green Chemistry-Synthesis Technology(GCTKF2014006)of Key Laboratory of High Efficient Processing of Bamboo of Zhejiang Province(2016)of State Key Laboratory of Chemical Resource Engineering,Beijing University of Chemical Technology(CRE-2016-C-303)~~
文摘The use of H2SO4‐,HCl‐,H3PO4‐,and CH3COOH‐activated montmorillonite(Mt)and WOx/H3PO4‐activated Mt as catalysts for the gas‐phase dehydration of glycerol was investigated.The WOx/H3PO4‐activated Mt catalysts were prepared by an impregnation method using H3PO4‐activated Mt(Mt‐P)as the support.The catalysts were characterized using powder X‐ray diffraction,Fourier‐transform infrared spectroscopy,N2adsorption‐desorption,diffuse reflectance ultraviolet‐visible spectroscopy,temperature‐programmed desorption of NH3,and thermogravimetric analysis.The acid activation of Mt and WOx loaded on Mt‐P affected the strength and number of acid sites arising from H+exchange,the leaching of octahedral Al3+cations from Mt octahedral sheets,and the types of WOx(2.7≤x≤3)species(i.e.,isolated WO4/WO6‐containing clusters,two‐dimensional[WO6]polytungstates,or three‐dimensional WO3crystals).The strong acid sites were weakened,and the weak and medium acid sites were strengthened when the W loading on Mt‐P was12wt%(12%W/Mt‐P).The12%W/Mt‐P catalyst showed the highest catalytic activity.It gave a glycerol conversion of89.6%and an acrolein selectivity of81.8%at320°C.Coke deposition on the surface of the catalyst led to deactivation.
基金the Razi University Research Council for support of this work
文摘A facile approach to the preparation of a novel magnetically separable H_5PMo_(10)V_2O_(40)/Fe_3O_4/g-C_3N_4(PMoV/Fe_3O_4/g-C_3N_4) nanocomposite by chemical impregnation is demonstrated.The prepared nanocomposite was characterized and its acidity was measured by potentiometric titration.PMoV/Fe_3O_4/g-C_3N_4 showed high catalytic activity in the selective oxidative desulfurization of sulfides to their corresponding sulfoxides or sulfones.The catalytic oxidation of a dibenzothiophene(DBT)-containing model oil and that of real oil were also studied under optimized conditions.In addition,the effects of various nitrogen compounds,as well as the use of one- and two-ring aromatic hydrocarbons as co-solvents,on the catalytic removal of sulfur from DBT were investigated.The catalyst was easily separated and could be recovered from the reaction mixture by using an external magnetic field.Additionally,the remaining reactants could be separated from the products by simple decantation if an appropriate solvent was chosen for the extraction.The advantages of this nanocatalyst are its high catalytic activity and reusability;it can be used at least four times without considerable loss of activity.
基金This work was supported by the National Natural Science Foundation of China(No.30170736)China National 863'High-tech Program(Grant No.2004AA603220).
文摘Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) plays important roles in various cellular processes. A cytosolic GAPDH encoding gene (gpd) of Gracilaria/Gracilariopsis lemaneiformis was cloned and characterized. Deduced amino acid sequence of the enzyme of G. lemaneiformis had high homology with those of seven red algae. The 5'-untranslated regions of the GAPDHs encoding genes of these red algae varied greatly. GAPDHs of these red algae shared the highly conserved glyceraldehyde 3-phosphate dehydrogenase active site ASCTTNCL. However, such active site of Cyanidium caldarium was different from those of the other six algae at the last two residues (CL to LF), thus the spatial structure of its GAPDH active center may be different from those of the other six. Phylogenetic analysis indicated that GAPDH of G. lemaneiformis might have undergone an evolution similar to those of Porphyra yezoensis, Chondrus crispus, and Gracilaria verrucosa. C. caldarium had a closer evolutionary relationship with Cyanidioschyzon merolae than with Cyanidium sp. Virtual Northern blot analysis revealed that gpd of G. lemaneiformis expressed constitutively, which suggested that it might be house-keeping and could be adapted as an inner control in gene expression analysis of G. lemaneiformis.
基金Supported by the National Natural Science Foundation of China(No.41176113)the National Basic Research Program of China(973 Program)(No.2010CB126403)+1 种基金the Changjiang Scholars Program for Innovative Research Team in Universities(No.IRT0941)the Earmarked Fund for Modern Agro-Industry Technology Research System(No.nycytx-47)
文摘Accurate quantification of transcripts using quantitative real-time polymerase chain reaction (qPCR) depends on the identification of reliable reference genes for normalization. This study aimed to identify and validate seven reference genes, including actin-2 (ACT-2), elongation factor 1 alpha (EF-1α), elongation factor 1 beta (EF-1β), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin (UBQ), β-tubulin (β-TUB), and 18 S ribosomal RNA, from Crassostrea angulata, a valuable marine bivalve cultured worldwide. Transcript levels of the candidate reference genes were examined using qPCR analysis and showed differential expression patterns in the mantle, gill, adductor muscle, labial palp, visceral mass, hemolymph and gonad tissues. Quantitative data were analyzed using the geNorm software to assess the expression stability of the candidate reference genes, revealing that β-TUB and UBQ were the most stable genes. The commonly used GAPDH and 18S rRNA showed low stability, making them unsuitable candidates in this system. The expression pattern of the G protein β-subunit gene (Gβ) across tissue types was also examined and normalized to the expression of each or both of UBQ andβ-TUB as internal controls. This revealed consistent trends with all three normalization approaches, thus validating the reliability of UBQ and β-TUB as optimal internal controls. The study provides the first validated reference genes for accurate data normalization in transcript profiling in Crassostrea angulata, which will be indispensable for further fimetional genomics studies in this economically valuable marine bivalve.
文摘An experiment was performed to determine the dietary phosphorus requirement of the young abalone, Haliotis discus hannai. Five semi-purified diets were formulated to provide a series of graded levels of dietary total phosphorus (0.23%-1.98) from monobasic potassium phosphate (KH2PO4). The brown alga, Laminaria japonica, was used as a control diet. Similar size abalone were distributed in a single-pass, flow-through system using a completely randomized design with six treatments and three replicates each treatment. The abalone were hand-fed to satiation with appropriate diets in excess, once daily at 17:00. The feeding trial was run for 120-d. Survival rate and soft-body to shell ratio (SB/S) were constantly maintained regardless of dietary treatment. However, the weight gain rate (WGR), daily increment in shell length (DISL), muscle RNA to DNA ratio (RNA/DNA), carcass levels of lipid and protein, soft-body alkaline phosphatase (SBAKP), and phosphorus concentrations of whole body (WB) and soft body (SB) were significantly (ANOVA, P<0.05) affected by the dietary phosphorus level. The dietary phosphorus requirements of the abalone were evaluated from the WGR, DISL, and RNA/DNA ratio respectively, by using second-order polynomial regression analysis. Based on these criteria, about 1.0%-1.2% total dietary phosphorus, i.e. 0.9%-1.1% dietary available phosphorus is recommended for the maximum growth of the abalone.
基金the National Natural Science Foundation of China(No.21266022)the National High Technology Research and Development Program 863(2012AA101800-03+4 种基金2012AA02120562012AA021704)the International Cooperation of Jiangxi Province(No.20101208)the International Science & Technology Cooperation Program of China(No.2010DFB63750)the Natural Science Foundation of Jiangxi Province(No.2008GZH0047)
文摘The carboxyl terminal of sodium oleate has a stronger polarity than that of oleic acid;this terminal is more likely to be dipole polarized and ionically conductive in a microwave field.Sodium oleate was used as the model compound to study the decarboxylation of oleic acid leading to hydrocarbon formation via microwave-assisted pyrolysis technology.The pyrolysis gas,liquid,and solid products were precisely analyzed to deduce the mechanism for decarboxylation of sodium oleate.Microwave energy was able to selectively heat the carboxyl terminal of sodium oleate.During decarboxylation,the double bond in the long hydrocarbon chain formed a p-πconjugated system with the carbanion intermediate.The resulting p-πconjugated system was more stable and beneficial to the pyrolysis reaction(decarboxylation,terminal allylation,isomerization,and aromatization).The physical properties of pyrolysis liquid were generally similar to those of diesel fuel,thereby demonstrating the possible use of microwaves for controlling the decarboxylation of sodium oleate in order to manufacture renewable hydrocarbon fuels.
基金Supported by the National Natural Science Foundation of China(21676288)
文摘The purpose of this study is to increase acrolein yield and capability of coking resistance in the reaction of glycerol dehydration to acrolein by assembling metal phosphate supported on HZSM-5 catalyst. The as-prepared catalysts were characterized by XRD, SEM, EDS, BET, NH_3-TPD, CO_2-TPD and Py-IR techniques. It was found that metal phosphate species were incorporated into the porous structure of HZSM-5 zeolites, thus influencing the surface and textural physico-chemical properties of the supporters. The alkaline-treated HZSM-5 catalyst promoted the dispersion of phosphate species on the carriers. Moreover, the amount of strong acidity was tremendously improved by adding the different metal hydrophosphates and the catalysts show high catalytic activity. In this present work, the Sn1/4 H_2PO_4/HZSM-5 catalyst exhibited good performance in the catalytic activity and coking resistant ability, which resulted in a high acrolein yield of 83% initially and acrolein yield of 68% after 30 h. The acidity, especially the ratio of strong to weak acid, plays an important role in promoting acrolein yield and stability simultaneously.
文摘Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), initially identified as a glycolytic enzyme and considered as a housekeeping gene, is widely used as an internal control in experiments on proteins, mRNA, and DNA. However, emerging evidence indicates that GAPDH is implicated in diverse functions independent of its role in energy metabolism; the expression status of GAPDH is also deregulated in various cancer cells. One of the most common effects of GAPDH is its inconsistent role in the determination of cancer cell fate. Furthermore, studies have described GAPDH as a regulator of cell death; other studies have suggested that GAPDH participates in tumor progression and serves as a new therapeutic target. However, related regulatory mechanisms of its numerous cellular functions and deregulated expression levels remain unclear. GAPDH is tightly regulated at transcriptional and pnsttranscriptional levels, which are involved in the regulation of diverse GAPDH functions. Several cancer-related factors, such as insulin, hypoxia inducible factor-1 (HIF-1), p53, nitric oxide (NO), and acetylated histone, not only modulate GAPDH gene expression but also affect protein functions via common pathways. Moreover, posttranslational modifications (PTMs) occurring in GAPDH in cancer cells result in new activities unrelated to the original glycnlytic function of GAPDH. In this review, recent findings related to GAPDH transcriptional regulation and PTMs are summarized. Mechanisms and pathways involved in GAPDH regulation and its different roles in cancer cells are also described.
文摘The removal of acid compounds (naphthenates) from acidic oil with ionic liquids was systematically investigated. [BMIM]Br-AlCl3 was used to investigate the effect on deacidification of oil. Experimental results showed that at a temperature of 323K with a molar ratio of AlCl3 to [BM1M]Br-AlCl3 of 0.2, and a mass ratio of IL to white oil of 4%, the deacidification rate could reach 75.9%. And a reaction time of 4 h was sufficient to achieve the goal. The study on reproducibility of catalytic performance of [BMIM]Br-AlCl3 showed the possibility of using the ionic liquid in the continuous catalytic reaction.
文摘The method for refining the straight-run diesel fuel was studied in laboratory scale in order to make the acid number of diesel fraction comply with the standard while removing the naphthenic acids contained in diesel without causing environmental pollution. After comparing the effect of refining using three solvents, the isopropyl alcohol-HOA was specified as the best solvent. Meanwhile, the relationship between the acid number of diesel fraction and the amount of solvent used and the relationship between the concentration of solvent and temperature and the stability of diesel in terms of its acid number were also investigated. Experimental results had shown that when the mass fraction of the HOA-IPA solvent was 20% at a dosage of 17 mL of the solvent and a temperature of 30℃, the acid number of the refined diesel fraction was 0.015 mg KOH/g with a good stability of acidity in the diesel fraction.
基金Supported by the National Natural Science Foundation of China(Nos.41476140,41306151,41676157,41506172)the Strategic Leading Science and Technology Projects of Chinese Academy of Sciences(No.XDA11020404)+1 种基金the China Postdoctoral Science Foundation(No.2015M582153)the Science and Technology Plan of Jiangsu Province(No.BE2016330)
文摘Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here, the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then, we used geNorm, a common statistical algorithm, to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while tubulin, glyceraldehyde- 3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification, we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress, and that a-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.
文摘The aim of the study was to evaluate the impact of organic selenium supplementation (selenium yeast) vs. inorganic selenium (selenium selenite) on the expression of selected defensin genes in milk somatic cells (MSC) of dairy cows. Sixteen Polish Holstein-Friesian cows in second parity, with similar milk yield and milk composition were randomly divided into two equal groups. At the beginning of experiment, the animals were in the middle of their lactation curve (150, standard deviation (SD) = 26 d) and this experiment lasted 90 d. The basic diet, for both groups, consisted of corn silage, wilted grass silage and concentrates. The diet of control group was supplemented with commercial mineral and vitamin mixture with inorganic selenium and the for the experimental group daily selenium need was covered by the addition of Se-yeast (6 mg Se/cow/d). Milk samples were collected three times during experiment (before experiment; on day 55; after the end the experiment). Total RNA was isolated from milk somatic cells and the levels of transcripts of bovine β1-defensin (DEFB1), β4-defensin (BNBD4), β5-defensin (BNBDS), β10-defensin (BNBD10) and lingual antimicrobial peptide (LAP) were measured with real time PCR, using glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a reference gene. The expressions of almost all studied genes were influenced by environmental conditions (very low and negative temperature). However, BNBD4 (P 〈 0.05) and LAP (P ≤ 0.001 ) genes were influenced by selenium supplementation but in opposite ways, depending on the form of Se. These findings support that selenium is an important factor affecting the mRNA level in MSC, but the effect of the form of selenium might depend on genes.