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1.
不同植茶年限茶园土壤pH缓冲容量 总被引:6,自引:0,他引:6
为探明长期植茶对土壤pH缓冲容量(pHBC)的影响,以安徽郎溪和祁门茶园为研究对象,研究了连续植茶10、15、20、25、30年的茶园土壤酸碱缓冲容量的变化及其影响因素.结果表明: 酸碱滴定法适用于茶园土壤pHBC的测定,酸碱加入量与pH值在酸碱滴定曲线的特定突跃段(pH4.0~6.0)呈近似直线关系,可通过线性拟合方程计算pHBC.两地茶园土壤的pHBC随着植茶年限的增加均呈下降趋势,郎溪茶园和祁门茶园土壤pHBC的下降速率分别为0.10和0.06 mmol·kg-1·a-1.茶园土壤pHBC与阳离子交换量、土壤有机质、盐基饱和度、物理性质黏粒含量呈显著正相关,而与交换性酸总量及交换氢含量呈显著负相关. 相似文献
2.
Sophie Richier Eric P. Achterberg Matthew P. Humphreys Alex J. Poulton David J. Suggett Toby Tyrrell Christopher Mark Moore 《Global Change Biology》2018,24(9):4438-4452
Accumulation of anthropogenic CO2 is significantly altering ocean chemistry. A range of biological impacts resulting from this oceanic CO2 accumulation are emerging, however, the mechanisms responsible for observed differential susceptibility between organisms and across environmental settings remain obscure. A primary consequence of increased oceanic CO2 uptake is a decrease in the carbonate system buffer capacity, which characterizes the system's chemical resilience to changes in CO2, generating the potential for enhanced variability in pCO2 and the concentration of carbonate [], bicarbonate [], and protons [H+] in the future ocean. We conducted a meta‐analysis of 17 shipboard manipulation experiments performed across three distinct geographical regions that encompassed a wide range of environmental conditions from European temperate seas to Arctic and Southern oceans. These data demonstrated a correlation between the magnitude of natural phytoplankton community biological responses to short‐term CO2 changes and variability in the local buffer capacity across ocean basin scales. Specifically, short‐term suppression of small phytoplankton (<10 μm) net growth rates were consistently observed under enhanced pCO2 within experiments performed in regions with higher ambient buffer capacity. The results further highlight the relevance of phytoplankton cell size for the impacts of enhanced pCO2 in both the modern and future ocean. Specifically, cell size‐related acclimation and adaptation to regional environmental variability, as characterized by buffer capacity, likely influences interactions between primary producers and carbonate chemistry over a range of spatio‐temporal scales. 相似文献
3.
Controlling pH is essential for a variety of biopharmaceutical process steps. The chemical stability of biologics such as monoclonal antibodies is pH‐dependent and slightly acidic conditions are favorable for stability in a number of cases. Since control of pH is widely provided by added buffer salts, the current study summarizes the buffer characteristics of acetate, citrate, histidine, succinate, and phosphate buffers. Experimentally derived values largely coincide with values calculated from a model that had been proposed in 1922 by van Slyke. As high concentrated protein formulations become more and more prevalent for biologics, the self‐buffering potential of proteins becomes of relevance. The current study provides information on buffer characteristics for pH ranges down to 4.0 and up to 8.0 and shows that a monoclonal antibody at 50 mg/mL exhibits similar buffer capacity as 6 mM citrate or 14 mM histidine (pH 5.0–6.0). Buffer capacity of antibody solutions scales linearly with protein concentration up to more than 200 mg/mL. At a protein concentration of 220 mg/mL, the buffer capacity resembles the buffer capacity of 30 mM citrate or 50 mM histidine (pH 5.0–6.0). The buffer capacity of monoclonal antibodies is practically identical at the process relevant temperatures 5, 25, and 40°C. Changes in ionic strength of ΔI=0.15, in contrast, can alter the buffer capacity up to 35%. In conclusion, due to efficient self‐buffering by antibodies in the pH range of favored chemical stability, conventional buffer excipients could be dispensable for pH stabilization of high concentrated protein solutions. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29: 480–492, 2013 相似文献
4.
G. M. Rodchenkov A. N. Vedenin V. P. Uralets V. A. Semenov 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1991,565(1-2)
Human urinary metabolites of the synthetic corticosteroids prednisone and prednisolone were detected in the course of gas chromatographic steroid profiling as methoxime-trimethylsilyl derivatives. Metabolites were provisionaly identified by combined gas chromatography—mass spectrometry. The major metabolites were 11-keto/11-hydroxy conversion products, 20-hydroxy and 4,5-dihydro analogues of the parent drugs. Cortisone, 6-hydroxy and fully saturated A-ring compounds were minor metabolites. Retention indices and mass spectral data are presented. 相似文献
5.
To realize coenzyme regeneration in the reduction of haloketones, a codon-optimized gene Sygdh encoding glucose 1-dehydrogenase (SyGDH) was synthesized based on the putative GDH gene sequence (Ta0897) in Thermoplasma acidophilum genomic DNA, and expressed in E. coli BL21(DE3). Recombinant SyGDH was purified to homogeneity by affinity chromatography with the specific activity of 86.3 U/mg protein towards D-glucose at the optimum pH and temperature of 7.5 and 40 °C. It was highly stable in a pH range of 4.5–8.0 and at 60 °C or below, and resistant to various organic solvents. The Km and catalytic efficiency (kcat/Km) of SyGDH towards NADP+ were 0.67 mM and 104.0 mM−1 s−1, respectively, while those towards NAD+ were 157.9 mM and 0.64 mM−1 s−1, suggesting that it preferred NADP+ as coenzyme to NAD+. Additionally, using whole cells of E. coli/Sygdh-Sys1, coexpressing SyGDH and carbonyl reductase (SyS1), as the biocatalyst, the asymmetric reduction of 60 mM m-chlorophenacyl chloride coupled with the regeneration of NADPH in situ was conducted in DMSO/phosphate buffer (2:8, v/v) system, producing (R)-2-chloro-1-(3-chlorophenyl)ethanol with over 99.9% eep and 99.2% yield. Similarly, the reduction of 40 mM α-bromoacetophenone in n-hexane/buffer (6:4, v/v) biphasic system produced (S)-2-bromo-1-phenylethanol with over 99.9% eep and 98.3% yield. 相似文献
6.
Sigrid Dengel Peter E. Levy John Grace Stephanie K. Jones Ute M. Skiba 《Global Change Biology》2011,17(12):3524-3533
Methane (CH4) is an important greenhouse gas, contributing 0.4–0.5 W m?2 to global warming. Methane emissions originate from several sources, including wetlands, rice paddies, termites and ruminating animals. Previous measurements of methane flux from farm animals have been carried out on animals in unnatural conditions, in laboratory chambers or fitted with cumbersome masks. This study introduces eddy covariance measurements of CH4, using the newly developed LI‐COR LI‐7700 open‐path methane analyser, to measure field‐scale fluxes from sheep grazing freely on pasture. Under summer conditions, fluxes of methane in the morning averaged 30 nmol m?2 s?1, whereas those in the afternoon were above 100 nmol m?2 s?1, and were roughly two orders of magnitude larger than the small methane emissions from the soil. Methane emissions showed no clear relationship with air temperature or photosynthetically active radiation, but some diurnal pattern was apparent, probably linked to sheep grazing behaviour and metabolism. Over the measurement period (days 60–277, year 2010), cumulative methane fluxes were 0.34 mol CH4 m?2, equating to 134.3 g CO2 equivalents m?2. By comparison, a carbon dioxide (CO2) sink of 819 g CO2 equivalents m?2 was measured over the same period, but it is likely that much of this would be released back to the atmosphere during the winter or as off‐site losses (through microbial and animal respiration). By dividing methane fluxes by the number of sheep in the field each day, we calculated CH4 emissions per head of livestock as 7.4 kg CH4 sheep?1 yr?1, close to the published IPCC emission factor of 8 kg CH4 sheep?1 yr?1. 相似文献
7.
8.
The capacity of soils to preserve organic C and N by their association with clay and silt particles 总被引:53,自引:3,他引:53
Jan Hassink 《Plant and Soil》1997,191(1):77-87
Although it has been recognized that the adsorption of organics to clay and silt particles is an important determinant of the stability of organic matter in soils, no attempts have been made to quantify the amounts of C and N that can be preserved in this way in different soils. Our hypothesis is that the amounts of C and N that can be associated with clay and silt particles is limited. This study quantifies the relationships between soil texture and the maximum amounts of C and N that can be preserved in the soil by their association with clay and silt particles. To estimate the maximum amounts of C and N that can be associated with clay and silt particles we compared the amounts of clay- and silt-associated C and N in Dutch grassland soils with corresponding Dutch arable soils. Secondly, we compared the amounts of clay- and silt-associated C and N in the Dutch soils with clay and silt-associated C and N in uncultivated soils of temperate and tropical regions.We observed that although the Dutch arable soils contained less C and N than the corresponding grassland soils, the amounts of C and N associated with clay and silt particles was the same indicating that the amounts of C and N that can become associated with this fraction had reached a maximum. We also observed close positive relationships between the proportion of primary particles < 20 m in a soil and the amounts of C and N that were associated with this fraction in the top 10 cm of soils from both temperate and tropical regions. The observed relationships were assumed to estimate the capacity of a soil to preserve C and N by their association with clay and silt particles. The observed relationships did not seem to be affected by the dominant type of clay mineral. The only exception were Australian soils, which had on average more than two times lower amounts of C and N associated with clay and silt particles than other soils. This was probably due to the combination of low precipitation and high temperature leading to low inputs of organic C and N.The amount of C and N in the fraction > 20 m was not correlated with soil texture. Cultivation decreased the amount of C and N in the fraction > 20 m to a greater extent than in the fraction < 20 m, indicating that C and N associated with the fraction < 20 m is better protected against decomposition.The finding of a given soil having a maximum capacity to preserve organic C and N will improve our estimations of the amounts of C and N that can become stabilized in soils. It has important consequences for the contribution of different soils to serve as a sink or source for C and N in the long term. 相似文献
9.
Nian Wu Jonathan V. Sweedler Mei Lin 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1994,654(2)
The four major bilirubin species in serum are separated by capillary electrophoresis and detected using laser-induced fluorescence detection. The optimum buffer system consists of 40 mM sodium dodecyl sulfate (SDS)—0.012 mM bovine serum albumin (BSA). The use of the SDS—BSA mixture in the mobile phase allows for the separation of four major bilirubin species at physiological pH with untreated capillaries. The results show that the use of BSA as a run buffer modifier in SDS solution improves separation efficiency and increases sample solubility via pH changes of the run buffer. The limits of detection for the bilirubin species using laser-induced fluorescence are between 30 and 150 nM, depending on the bilirubin species; not only is this approximately two orders of magnitude lower than with visible-light absorption methods, it allows the bilirubin species in normal sera to be quantitatively measured without sample pretreatment. 相似文献
10.
Meneses SR Marques KL Pires CK Santos JL Fernandes E Lima JL Zagatto EA 《Analytical biochemistry》2005,345(1):90-95
An automated flow-based procedure for assessment of total antioxidant capacity was developed. It involved a multipumping flow system, a recent approach to flow analysis, and exploited the ability of selected compounds to inhibit the chemiluminescence reactions of luminol or lucigenin with hydrogen peroxide. The system included several discretely actuated solenoid micropumps as the only active components of the flow manifold. This enabled the reproducible insertion and efficient mixing of very low volumes of sample and reagents as well as the transportation of the sample zone toward a flow-through luminometer, where the chemiluminometric response was monitored. With luminol as the chemiluminogenic reagent, linearity of the analytical curves was noted up to 3.2x10(-4), 1.1x10(-3), and 8.8x10(-8) molL-1 for Trolox, ascorbic acid, and resveratrol, respectively. With lucigenin, linear calibration plots up to 2x10(-5) molL-1 of Trolox and 5.7x10(-5)molL-1 of ascorbic acid were obtained. As favorable analytical figures of merit, the measurement precision (RSD typically between 0.2 and 2.0%, n=10), low operational costs, low reagent consumption, sampling rate (160 and 70 h-1), and versatility should be highlighted. The proposed system can be used in distinct analytical circumstances without requiring physical reconfiguration. 相似文献
11.
Characterization of Botrytis–plant interactions using PathTrack©—an automated system for dynamic analysis of disease development 下载免费PDF全文
Elad Eizner Mordechi Ronen Yonatan Gur Assaf Gavish Wenjun Zhu Amir Sharon 《Molecular Plant Pathology》2017,18(4):503-512
The measurement of disease development is integral in studies on plant–microbe interactions. To address the need for a dynamic and quantitative disease evaluation, we developed PathTrack©, and used it to analyse the interaction of plants with Botrytis cinerea. PathTrack© is composed of an infection chamber, a photography unit and software that produces video files and numerical values of disease progression. We identified a previously unrecognized infection stage and determined numerical parameters of pathogenic development. Using these parameters, we identified differences in disease dynamics between seemingly similar B. cinerea pathogenicity mutants, and revealed new details on plant susceptibility to the fungus. We showed that the difference between the lesion expansion rate on leaves and colony spreading rate on artificial medium reflects the levels of the plant immune system, suggesting that this parameter can be used to quantify plant defence. Our results shed new light and reveal new details of the interaction between the model necrotrophic pathogen B. cinerea and plants. The concept that we present is universal and may be applied to facilitate the study of various types of plant–pathogen association. 相似文献
12.
C. Loret B. Chaufer B. Sebille M. Hanselin Y. Blain A. Le Hir 《International journal of biological macromolecules》1988,10(6):366-372
Gelatin samples obtained by chemical modification (succinylation) are studied by SEC on silica based chromatographic supports. The influence of the pH of eluent mixtures (potassium phosphate added to NaCl) in the range 7-3.3 shows that the void volume peak (VVP) is lowered or even vanishes at pH 3.3 with the 3000 SW (TSK) gel. A process using an ultrasound treatment before injection is reported in order to determine accurately the molecular parameters of gelatin onto TSK gel with a minimal VV P. This peak is attributed to molecular aggregation of a part of the modified gelatin. After disaggregation by ultrasound or heat treatment the results are in good accordance with those obtained by other methods. It is demonstrated that with proteins and dextrans the TSK 3000 SW gel does not agree with the universal calibration curve (log[ν] ·
versus Kd as reported previously. A single calibration curve is obtained when the Stokes radius is plotted versus Kd. Gelatin fractions are eluted at pH 7 close to this calibration curve. This plot shows that gelatin fractions at pH 3.3 are not eluted by a pure size exclusion mechanism on 3000 SW gel. It is concluded that hydrophobic interactions between fractions of gelatin and the gel explain the high retention of these samples. 相似文献
13.
J. E. Butler E. P. Lü P. Navarro B. Christiansen 《Journal of molecular recognition : JMR》1997,10(1):36-51
Polydimethylsiloxane (PEP) is widely used in medical prostheses and therefore is in contact with plasma and secretory proteins. Two pair of globular proteins, lactoferrin (Lf) and transferrin (Trf), and bovine IgG1 and IgG2a, which differ substantially between pair members in their pl, were used to study the interaction of a PEP widely used in breast implants and soluble protein. Studies were done using iodinated proteins over a concentration range that resulted in an apparent protein monolayer. Secondary incubations with dilute protein solutions were needed to form the monolayer on PEP, possibly as a consequence of micro air bubbles trapped on its highly textured surface as shown by atomic force microscopy. Immunoassay quality polystyrene microtiter wells were used as controls. Adsorption studies were routinely performed at pH 4, 7 and 10 and at ionic strengths corresponding to 0.95, 9.5 and 90.0 mS. The protein capture capacity (PCC) of PEP for Lf and Trf was optimal at physiological pH and ionic strength and comparable under these conditions to that of Immulon 2 (Imm 2) microtiter wells. While increasing the ionic strength and pH further increases the PCC of Imm 2 for Lf and Trf, this markedly lowered the PCC of PEP for these proteins suggesting that initial polar interactions may precede subsequent hydrophobic bonding to PEP. This was tested using a hydrophilic variant of PEP, which when tested in a 90.0 mS buffer, showed a >five-fold lower PCC at neutral and alkaline pH. The greatly reduced PCC of the hydrophilic variant might also suggest that hydrophilic variants of silicone would be more biocompatible than those currently used. The PCC of PEP for the IgGs was less than that of Imm 2 but still optimal at physiological conditions. Consistent with the data on Lf/Trf, PCC progressively decreased with increasing ionic strength at alkaline pH. Differences in pl between the protein pairs had only a marginal effect on the PCC of PEP. Monolayer adsorption on both PEP and Imm 2 was slowly reversible and greater in the presence of free ligand (<2% in 16 h) suggesting that the process follows Mass Law principles. However, even in the presence of non-ionic detergent and free ligand, 85–90% remained bound on either surface. Thus, desorption of proteins in the monolayer should not complicate subsequent immunochemical studies conducted on adsorbed monolayers. © 1997 John Wiley & Sons, Ltd. 相似文献
14.
A wild-type strain of Candida albicans (S1, ATCC 10261) was used to obtain stable auxotrophic colony morphological mutants (mutant M5 producing only true hyphae and mutant M2 containing 90 % blastospores and 10 % pseudohyphae) by induced mutagenesis. A hybrid was produced by somatic hybridization between these 2 mutants. Out of the isolated 10 clones, 2 stable hybrid clones were chosen and characterized: clone VI. 1M produced rough colonies containing a new, extended cell type (never observed in natural isolates), exhibited unipolar budding, did not form a germ tube, and possessed 12 chromosomal bands. All other features (antifungal and stress sensitivity, adhesion ability, pathogenicity, and isoenzyme and RAPD patterns) were similar to those of mycelial mutant M5. In contrast, the characteristics of clone VI.9S were similar to those of morphological mutant M2. 相似文献
15.
Escherichia coli alkaline phosphatase (AP) and human lysozyme (h-LYZ), which contain two and four disulfide bonds, respectively, were expressed in a cell-free protein synthesis system constructed from Spodoptera frugiperda 21 (Sf21) cells. AP was expressed in a soluble and active form using the insect cell-free system under non-reducing conditions, and h-LYZ was expressed in a soluble and active form under non-reducing conditions after addition of reduced glutathione (GSH), oxidized glutathione (GSSG), and protein disulfide isomerase (PDI). The in vitro synthesized proteins were purified by means of a Strep-tag attached to their C termini. Approximately 41 microg AP and 30 microg h-LYZ were obtained from 1 mL each of the reaction mixture. The efficiency of protein synthesis approached that measured under reducing conditions. Analysis of the disulfide bond arrangements by MALDI-TOF MS showed that disulfide linkages identical to those observed in the wild-type proteins were formed. 相似文献
16.
Nora B. Caberoy Yixiong Zhou Xiaoyu Jiang Gabriela Alvarado Wei Li 《Journal of molecular recognition : JMR》2010,23(1):74-83
Mutation in the tubby gene causes adult‐onset obesity, progressive retinal, and cochlear degeneration with unknown mechanism. In contrast, mutations in tubby‐like protein 1 (Tulp1), whose C‐terminus is highly homologous to tubby, only lead to retinal degeneration. We speculate that their diverse N‐terminus may define their distinct disease profile. To elucidate the binding partners of tubby, we used tubby N‐terminus (tubby‐N) as bait to identify unknown binding proteins with open‐reading‐frame (ORF) phage display. T7 phage display was engineered with three improvements: high‐quality ORF phage display cDNA library, specific phage elution by protease cleavage, and dual phage display for sensitive high throughput screening. The new system is capable of identifying unknown bait‐binding proteins in as fast as ~4–7 days. While phage display with conventional cDNA libraries identifies high percentage of out‐of‐frame unnatural short peptides, all 28 tubby‐N‐binding clones identified by ORF phage display were ORFs. They encode 16 proteins, including 8 nuclear proteins. Fourteen proteins were analyzed by yeast two‐hybrid assay and protein pull‐down assay with ten of them independently verified. Comparative binding analyses revealed several proteins binding to both tubby and Tulp1 as well as one tubby‐specific binding protein. These data suggest that tubby‐N is capable of interacting with multiple nuclear and cytoplasmic protein binding partners. These results demonstrated that the newly‐engineered ORF phage display is a powerful technology to identify unknown protein–protein interactions. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
17.
Jasbir Chauhan Andr Darbre 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1980,183(4)
The determination of urinary homovanillic, isohomovanillic and vanillylmandelic acids as their trifluoroacetylhexafluoroisopropyl ester derivatives by glass capillary gas—liquid chromatography has been studied. It was shown that even with high column efficiencies a single peak—single compound relationship could not be assumed and for reliable quantitation it was necessary to check determinations with a second gas—liquid chromatography column. 相似文献
18.
Spyros Petrakis Martin H. Schaefer Erich E. Wanker Miguel A. Andrade‐Navarro 《BioEssays : news and reviews in molecular, cellular and developmental biology》2013,35(6):503-507
Polyglutamine (polyQ) diseases are genetically inherited neurodegenerative disorders. They are caused by mutations that result in polyQ expansions of particular proteins. Mutant proteins form intranuclear aggregates, induce cytotoxicity and cause neuronal cell death. Protein interaction data suggest that polyQ regions modulate interactions between coiled‐coil (CC) domains. In the case of the polyQ disease spinocerebellar ataxia type‐1 (SCA1), interacting proteins with CC domains further enhance aggregation and toxicity of mutant ataxin‐1 (ATXN1). Here, we suggest that CC partners interacting with the polyQ region of a mutant protein, increase its aggregation while partners that interact with a different region reduce the formation of aggregates. Computational analysis of genetic screens revealed that CC‐rich proteins are highly enriched among genes that enhance pathogenicity of polyQ proteins, supporting our hypothesis. We therefore suggest that blocking interactions between mutant polyQ proteins and their CC partners might constitute a promising preventive strategy against neurodegeneration. 相似文献
19.
Biomass carbon density and carbon sequestration capacity in seven typical forest types of the Xiaoxing’an Mountains,China 总被引:2,自引:0,他引:2 下载免费PDF全文
森林生物碳储量作为森林生态系统碳库的重要组成部分,在全球碳循环中发挥着重要作用。以小兴安岭7种典型林型为研究对象,通过外业样地调查与室内实验分析相结合的方法,从林分尺度对林分生物量与碳密度进行计量,分析了林分生物碳储量的空间分配格局,并对林分年固碳能力与碳汇潜力进行了探讨。结果表明:小兴安岭不同林型从幼龄林到成熟林的乔木层碳密度增长速率为:蒙古栎(Quercus mongolica)林>兴安落叶松(Larix gmelinii)林>云冷杉(Picea-Abies)林>樟子松(Pinus sylvestris var.mongolica)林>山杨(Populus davidiana)林>红松(Pinus koraiensis)林>白桦(Betula platyphylla)林。7种典型林型不同龄组(幼龄林、中龄林、近熟林和成熟林)林分生物量碳密度分别为:红松林31.4、74.7、118.4和130.2 t·hm–2;兴安落叶松林28.9、44.3、74.2和113.3 t·hm–2;樟子松林22.8、52.0、71.1和92.6 t·hm–2;云冷杉林23.1、44.1、77.6和130.3 t·hm–2;白桦林18.8、35.3、66.6和88.5 t·hm–2;蒙古栎林25.0、20.0、47.5和68.9 t·hm–2;山杨林19.8、28.7、43.7和76.6 t·hm–2。红松林、兴安落叶松林、樟子松林和蒙古栎林在幼龄林时林分年固碳量较高,其他林型在成熟林时林分年固碳量较高。7种典型林型不同龄组的林分生物量碳密度均随林龄增长而增加,但不同林型的碳汇功能存在差异,同一林型不同林龄的生物量碳密度增幅差异也较大。林分年固碳量在0.4–2.8 t·hm–2之间,碳汇能力较强、碳汇潜力较大。尤其是小兴安岭目前林分质量较差,幼龄林和中龄林所占的比重较大,具有较大的碳汇潜力。研究结果可为森林经营管理及碳汇功能评价提供参考。 相似文献
20.
Characterization of in-frame proteins encoded by cvaA, an essential gene in the colicin V secretion system: CvaA* stabilizes CvaA to enhance secretion. 下载免费PDF全文
Colicin V (ColV), an antibacterial peptide toxin, uses a dedicated signal sequence-independent export system for its extracellular secretion in Escherichia coli. The products of at least three genes (a chromosomal tolC gene and two plasmid-born cvaA and cvaB genes) are involved in this process. To characterize the gene products, the cvaA gene was subcloned and expressed under the control of T7 RNA polymerase promoter. Two in-frame proteins, CvaA and CvaA*, were expressed and identified. DNA sequences predicted that both proteins have two potential translational initiation sites. N-terminal peptide sequencing showed that the translation of CvaA starts from a TTG, 11 amino acids upstream of the previously proposed ATG initiation site. CvaA* is translated from an upstream ATG. Expression of both CvaA and CvaA* was induced by the iron chelator 2,2'-dipyridyl, indicating that cvaA is negatively regulated at least partially by Fur. CvaA*-depleted cells were found to secrete less ColV, based on reduced activity in the supernatant, than did wild type, which was recovered by the addition of a plasmid producing CvaA*. Interestingly, CvaA*-depleted and wild-type cells had similar levels of intracellular ColV activity. Translational fusions showed that the syntheses of ColV and CvaA are not affected by CvaA* depletion. However, CvaA in CvaA*-depleted cells was less stable than that in wild-type cells, indicating that CvaA* may directly or indirectly affect the stability of CvaA. We conclude that CvaA* is not essential for ColV secretion but that it enhances the ColV secretion by stabilizing the CvaA protein. 相似文献