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为探索节旋藻藻蓝蛋白的生物合成机理,以钝顶节旋藻(Arthrospira platensisFACHB314)基因组DNA为模板克隆了藻蓝蛋白α亚基基因cpcA、催化脱辅基蛋白与色基结合的裂合酶基因cpcE和cpcF,以及催化色基合成的铁氧蛋白氧化还原酶基因pcyA;以Synechocystissp.PCC6803基因组DNA为模板克隆了亚铁血红素氧化酶基因hox1。然后分别将cpcA、cpcE和cpcF基因构建到质粒pACYCDuet-1中,将hox1和pcyA基因构建到质粒pET-24a(+)中,用电击法将二者共同转化E.coliBL21(DE3),经诱导表达得到具有荧光活性的节旋藻藻蓝蛋白α亚基。在590 nm激发波长下,荧光发射峰为637.8 nm,证实了节旋藻自身的裂合酶CpcE和CpcF能够催化色基PCB与藻蓝蛋白脱辅基结合产生有荧光活性的藻蓝蛋白α亚基,为表达有荧光活性的藻蓝蛋白提供理论和实验基础。 相似文献
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藻胆蛋白的研究概况(Ⅱ)——藻胆蛋白的结构及其光谱特性 总被引：15，自引：2，他引：13
１藻胆蛋白的一级结构与特异位点的修饰目前,已经对一些藻胆蛋白的一级结构进行了测定。测定的方法有两种,一种是直接测定,另一种是测定相应基因的核苷酸序列再反推其氨基酸序列。通过对一级结构的比较分析,发现藻胆蛋白在某些位点处的氨基酸残基相当保守,并且认为这些位点在保证色基的构象以及蛋白质分子稳定性方面具有重要的意义。在一级结构分析中,还发现了一个相当有意义的现象,即在所有已测定的藻胆蛋白分子中,其β亚基的７２位均为经过修饰的γＮ甲酰天冬酰胺残基,这是经蛋白质的转译后修饰而产生的,而且不论是原核藻类还是真核藻类的… 相似文献
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钝顶螺旋藻C-藻蓝蛋白和多管藻R-藻红蛋白的分离纯化及摩尔消光系数的测定 总被引：2，自引：1，他引：2
１９９３年１１月～１９９４年１月用简单的羟基磷灰石柱层析法从多管藻中分 离纯化了Ｒ－藻红蛋白和从钝顶螺旋藻中纯化了Ｃ－藻蓝蛋白，它们的纯度（指可见 光部分的最大吸收与 ２８０ｎｍ处吸收值之比）可分别达到 ６（Ｒ－藻红蛋白）和 ５． ５（Ｃ－ 藻蓝蛋白）。由于不同藻种的同种藻胆蛋白的摩尔消光系数不同，所以应用凯氏定 氮法并结合可见光的吸收值测定了上述两种藻胆蛋白的摩尔消光系数，钝顶螺旋 藻Ｃ－藻蓝蛋白为１．８５３ × １０６ｍｏｌ－１ｃｍ－１（６２０ｎｍ），多管藻Ｒ－藻红蛋白为１．７９６ × １０６ｍｏｌ－１ｃｍ－１（４９８ｎｍ），为藻胆蛋白的浓度测定提供了方便。 相似文献
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蓝隐藻藻蓝蛋白结构与功能稳定性研究 总被引：1，自引：0，他引：1
以蓝隐藻(Chroomonas placiodea)藻蓝蛋白 PC-645为材料,通过改变环境 pH 和尿素质量浓度,监测其变性和复性过程中特征荧光谱和吸收谱的动力学变化,以期了解隐藻藻蓝蛋白的色基和蛋白结构稳定性及其与功能的关系。结果表明, PC-645在很宽的 pH 范围和一定质量浓度的尿素中维持结构和功能的稳定,空间结构具有很强的柔性。pH诱导的 PC-645蛋白构象与功能变化可分为3个不同的区段。(1)稳定区(pH 3.5~7)：吸收和荧光光谱都比较稳定,显示蛋白质构象和功能在此区域都保持正常。(2)次稳定区(pH 7~10)：光吸收依然保持平稳,亚基内部的色基的状态和疏水微环境都没有改变,但荧光传递效率降低,可能是由亚基表面局部构象变化、解离(四级结构变化)或者色素基团间的空间距离变化引起。(3)不稳定区(pH<3.5和 pH>10),吸光度和荧光强度都呈快速下降,色基在近紫外和可见光区的吸收峰位变动,蛋白构象处于快速崩溃期。PC-645在酸性条件下的稳定性高于在碱性环境下,是与隐藻藻蓝蛋白所处的特殊环境及生理功能相适应的。 相似文献
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用N-甲基-N′-硝基-N-亚硝基胍（MNNG）对龙须菜配子体进行诱变处理，并筛选出色素突变体。又以真江蓠、细基江篱繁枝变型、龙须菜及其突变体为材料进行了藻胞蛋白的提取、纯化及藻胆蛋白光谱特性的研究。结果表明，江蓠属三个物种藻胆蛋白的含量有明显差异，龙须菜的野生型藻体与突变体之间藻胆蛋白含量的差异比不同物种之间的差异更为显著。江蓠属三个物种中藻胆蛋白的吸收光谱中以藻红蛋白（PE）变化最大：细基江蓠繁枝变型PE的吸收光谱有二峰一肩，属Ⅰ型R-PE，而真江蓠和龙须菜的PE的吸收光谱有三个峰，属Ⅱ型R－PE：龙须菜的不同突变型藻体的吸收光谱也有Ⅰ型和Ⅱ型R－PE之分。不同物种藻蓝蛋白也有R－PC和C－PC的差异，异藻蓝蛋白的吸收光谱及三种藻胆蛋白的荧光发射光谱也略有变化。并以光合进化的观点分析了本文的实验结果。 相似文献
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藻胆体是红、蓝藻特有的光合作用捕光天线复合物。别藻蓝蛋白(APC)是组成藻胆体高效能量传递的核心结构的主要成分。本文以基因重组的别藻蓝蛋白(rAPC)的单体和三聚体为材料,通过稳态光谱、圆二色光谱以及超快时间分辨光谱研究了rAPC的结构构象和能量传递过程。结果表明,rAPC在测试条件下能保持和天然APC一致的光谱特性和活性构象;rAPC单体组装成三聚体后,其α84PCB和β84PCB可以组成激子色素对,通过激子分裂提高三聚体的能量传递效率;超快时间分辨光谱结果显示,在rAPC三聚体中,能量从620 nm传至650 nm的时间为300～600 fs,同时存在着19 fs的激子态的电子退相干过程。这些结果为揭示藻胆体的高效能量传递机制提供了数据基础。 相似文献
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植物的光合器官通常含有叶绿素、类胡罗卜素和藻胆蛋白三类色素。藻胆蛋白主要包括藻红蛋白、藻蓝蛋白和变藻蓝蛋白。藻胆蛋白是水溶性的,这些蛋白质以共价键与发色团相连。R代表红藻纲,C代表蓝藻纲。 藻胆蛋白主要分布在蓝藻、红藻和隐藻中,是光合作用的辅助色素,当藻红蛋白吸收了光能后,将能量传递给藻蓝蛋白,然后经过变藻蓝蛋白传递给叶绿素a以进行光合作用。因而,藻胆蛋白是进行光合作用的主要光接受器。 相似文献
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The optical properties and distribution of dissolved organic matter in the surface waters of the Kara Sea and bays of Novaya Zemlya archipelago were studied during the 63th cruise of the R/V Akademik Mstislav Keldysh. The fluorescence of dissolved organic matter has been studied over wide excitation (230–550 nm) and emission (240–650 nm) wavelength ranges. Based on the results of fluorescence measurements, we propose a simple technique for estimating the relative content of humic compounds entering the Kara Sea shelf region with Ob and Yenisei river runoff. We have found that the blue shift parameters of the DOM fluorescence are Δ270–310 = 28 ± 2 nm and Δ355–310 = 29 ± 2 nm. The highest contents of humic compounds in surface waters were measured on the transect across the desalinated layer of the Kara Sea, near the continental slope on the transect along the St. Anna Trough, and in the area of Sedova, Oga and Tsivol’ki bays. Traces of labile terrigenous organic matter were found in the region of the Voronin Trough, in the bays of the Severny Island of Novaya Zemlya, as well as in some freshwater reservoirs and ice samples of the archipelago. We established a conservative distribution of dissolved organic matter, whose content in water varied from 1.25 to 8.55 mg/L. 相似文献
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高原湖泊溶解有机质的三维荧光光谱特性初步研究 总被引：9，自引：0，他引：9
提要近年来,荧光光谱技术被广泛应用于研究天然水体中溶解有机质(Dissolved Organic Matter,DOM)的物理化学特性。为了理解高原湖泊中DOM的组成、荧光光谱特性及其在湖泊水体中的垂直分布情况,作者利用荧光发射光谱、三维荧光光谱研究云贵高原湖泊红枫湖和百花湖中的DOM。结果显示,高原湖泊DOM主要表现为类富里酸荧光,包括可见区和紫外区两种类型的荧光峰,各种天然水体中都有报道的类蛋白荧光在红枫湖DOM中并不明显,而在百花湖DOM中则有较强的类蛋白荧光。溶解有机质所含三种类型荧光峰(PeakA:紫外区类富里酸荧光峰;PeakB:可见区类富里酸荧光峰;PeakC:类蛋白荧光峰)的荧光强度与溶解有机碳之间没有明显的线性相关关系,可能与高原湖泊水体pH值、DOM在湖泊不同深度由于受到光降解、微生物降解等作用存在差异等因素有关。与有机质结构和成熟度有关的荧光峰比值r(A,C)在1.40—2.09范围内,红枫湖南湖、百花湖DOM样品的r(A,C)值随着水体深度有下降的趋势,而红枫湖北湖DOM的r(A,C)值则随着水体深度有较大起伏,揭示了高原湖泊水体中DOM的结构以及分布情况存在差异。另外,r(A,C)值与pH之间具有良好的正相关关系,其相关系数红枫湖南湖DOM为0.95,百花湖两个采样点DOM分别为0.67、0.75,而红枫湖北湖DOM则显示出一定的负相关性(R2=0.45)。 相似文献
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《Deep Sea Research Part II: Topical Studies in Oceanography》1999,46(8-9):1769-1790
Scanning fluorescence spectroscopy was used to investigate the spatial and temporal variability in the fluorescence signature of phycoerythrin-containing organisms in the Arabian Sea during the early Northeast and early Southwest Monsoon (1994–1995). Phycoerythrin (PE) emission spectra were relatively invariant among all the samples collected on either cruise; the relatively symmetrical PE emission peaks showed maxima at wavelengths ranging from 563–572 nm. PE excitation spectra always showed either a strong shoulder or a peak at wavelengths absorbed maximally by phycourobilin (PUB) chromophores as well as a peak at wavelengths absorbed maximally by phycoerythrobilin (PEB) chromophores. Thus, the Arabian Sea appears to be different from the Black Sea or Gulf of Maine in that PUB-lacking forms of PE rarely, if ever, dominate the PE signal. Fluorescence excitation signatures differed in the relative excitation of PE emission by wavelengths absorbed by PUB (∼495 nm, ExPUB) and by wavelengths absorbed by PEB (∼550 nm, ExPEB); these were distinguished by having either very low (∼0.6), very high (∼1.8), or intermediate ExPUB:ExPEB ratios. The distribution of samples with different PE fluorescence signatures was investigated extensively during the early Southwest Monsoon, and communities characterized by the low ExPUB:ExPEB ratios were closely associated with cooler (24–27°C), fresher (35.7–36.25 psu) water influenced by coastal upwelling. In general, “ambient” surface water of the Arabian Sea during the early Southwest Monsoon was of intermediate temperature (27–29°C) and salinity (36.15–36.4 psu) and showed intermediate or high values for ExPUB:ExPEB. This suggests that the PE fluorescence signature can be used to follow the fate of upwelling-influenced water masses and the populations they transport. 相似文献
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Abstract. In this paper we present a one-year investigation on the fluorescence characteristics of released products of selected Adriatic seaweeds. The technique chosen was UV-fluorescence spectroscopy with the synchronous mode, which involves the stepping of the excitation and emission monochromators. The five species, of representative macrophytes, Cystoseira compressa, Dictyopreris polypodioides, Gracilaria bursapastoris, Pterocladia capillacea, and Ulva rigida, were chosen on the basis of their dominance in the plant communities typical for the area near Split - Central Adriatic. Our results suggest that macroalgal-related dissolved fluorescent matter can be categorized into two distinct groups of fluorescing compounds with the following excitation wavelengths: one from 320–360 nm, probably for compounds with 2 fused aromatic rings, and another from 360–420 nm, corresponding with 3 & 4 fused rings. The contribution of aromatics with 2 and with 3 & 4 rings was very similar in the brown algae Cysroseira compressa and Dictyopteris polypodioides, while in the red algae Gracilaria bursapastoris and Pterocludia capillacea as well in the green alga Ulva rigida the group with 2 rings dominated. 相似文献
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2007年夏季在东海舟山海域河口锋区开展了陆源溶解有机质的调查研究。测定了有色溶解有机质(CDOM)在激发波长370 nm/发射波长460 nm处的荧光强度和在λ=355 nm处的吸收系数,用于代表陆源CDOM浓度,并测定了荧光指数以指示CDOM来源。结果表明,CDOM的荧光值和紫外吸收系数之间呈显著正相关性,陆源CDOM浓度大体有向海方向降低的趋势,但是纵向上存在一些"突跃"现象。在舟山海域东北角不时观测到表层水体含有高浓度的CDOM,但变异性很大,推测可能该海区受到长江口羽状流的影响。在连续观测站发现陆源CDOM浓度在低平潮时往往比高平潮时要高。河海水在混合过程中CDOM浓度与盐度呈显著的线性负相关关系。在低盐度的悬沙锋区(S<24)CDOM浓度明显低于理论稀释值,而在较高盐度的羽状锋区,CDOM浓度接近于理论稀释值。在盐度为24~31范围内,大部分水样的荧光指数在1.50上下波动,表明其中CDOM来源以陆地来源为主;在较低盐度(S<24)的水样中荧光指数在1.70至1.90以上,表明CDOM以海洋来源为主,这与其陆源组分在高浊度的低盐度区存在显著的去除过程有关。研究表明,舟山海域水质存在着显著的变异性,与近岸羽状流密切相关,陆源溶解有机质的分布特征对此有较好的响应。 相似文献
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Phosphorus Near Edge X-ray Fluorescence Spectroscopy (P-NEXFS) data were collected on phosphorus containing phases including organic and inorganic compounds and minerals. Although phases containing P in the plus five oxidation state P(V) in a tetrahedral PO4 structure have similar primary fluorescence peak positions, the size, shape, and positions of secondary spectral features are diagnostic for different compounds and minerals. In particular, calcium phosphates exhibited a notable post-peak shoulder at 2154.5 eV, while oxidized iron phosphates had a distinctive pre-peak feature at 2148 eV. Polyphosphates have a broad secondary peak located approximately 2 eV higher in energy than a similar feature in phosphate esters and diesters. Compounds containing P(V) in structures other than PO4 tetrahedra such as phosphonates have a primary peak shifted about 1 eV lower than corresponding organo-phosphates. Organo-phosphates with P in the plus three oxidation state P(III) such as phosphines had primary fluorescence peaks shifted still further down in energy (2–3 eV). The substitution of aromatic carbon groups in close proximity to P structures in organic compounds generated both pre- and post-peak features as well as a number of secondary peaks. In addition, X-ray fluorescence mapping of P, Si, Al, Mg, and Na was conducted on a marine sediment sample with sub-micron spatial resolution. Phosphorus was heterogeneously distributed in the sample and not correlated on a broad scale with any other element examined. Much of the P present in the sample was located in small, 0.6–8 μm size, P-rich domains. Several P-rich regions were examined with P-NEXFS using a focused beam with 60 nm resolution and were found to consist of either calcium phosphate or polyphosphate phases. The presence of significant polyphosphate-dominated regions in a marine sediment sample supports the recent observations that such phases can play an important role in marine P cycling. The combination of fluorescence mapping and P-NEXFS data collection on fine particles provides a powerful new tool for environmental phosphorus studies. 相似文献