重组藻胆蛋白与高等植物类囊体膜之间能量传递的研究     

尚孟慧  臧晓南  林娇娇  李瑞  毕莹  徐晓婷 《中国海洋大学学报(自然科学版)》2021,(5)

为了探究藻胆蛋白与高等植物类囊体膜之间的光能传递规律,验证重组藻胆蛋白是否可以与高等植物类囊体膜之间的光能传递,本研究分别提取了重组藻红蛋白、重组藻蓝蛋白以及菠菜和韭菜的类囊体进行混合孵育,然后进行了全波长吸收光谱以及荧光光谱检测。结果显示:混合孵育时间为10~20 min有利于重组藻红蛋白和藻蓝蛋白与类囊体膜结合并进行光能传递,最适混合孵育时间随重组藻胆蛋白和类囊体的种类不同而稍有不同;重组藻红蛋白不能与菠菜或韭菜的类囊体膜直接发生光能传递,但是当体系中添加重组藻蓝蛋白的时候就可以;重组藻蓝蛋白可以与类囊体中的叶绿素a直接进行光能传递,使得叶绿素a的荧光发射峰值显著提升,当藻红蛋白和藻蓝蛋白的比例是1∶2时,光能传递效率较高。本研究为藻胆蛋白在构建新型光合系统中的应用提供实验依据。  相似文献   

倒立水母Cassiopea andromeda 的POU、Hox基因家族的多样性及系统发育分析          下载免费PDF全文

陈依婧  郑森林  彭胜蓝  林茂  付腾苇 《应用海洋学学报》2021,40(4):587-596

与其他钵水母相比,倒立水母具有独特行为：在生命周期的大部分时间里,它都在海底呈倒立附着、“睡眠”的状态。为探索与这一独特行为相关的遗传信息,本研究对安朵仙水母(Cassiopea andromeda)和同属于钵水母纲的海蜇(Rhopilema esculentum)、巴布亚硝水母(Mastigias papua)进行首次全基因组测序、拼接和注释,并重点分析了这3种钵水母与感觉功能、神经系统发育相关的重要转录调节因子Hox和POU基因家族的多样性与系统发育关系。遗传分析显示,刺胞动物门中Hox和POU基因家族具有明显的物种间差异性。对Hox基因的分析首次发现钵水母及水螅(Hydra vulgaris)的“前段Hox基因”产生了部分缺失,并进一步印证了刺胞动物不存在“中段Hox基因”的假说。安朵仙水母和海蜇具有相对全面的ParaHox基因种类,即GSX和XLOX/CDX基因,而巴布亚硝水母只有GSX基因。在POU基因多样性方面,安朵仙水母、海蜇、星状海葵(Nematostella vectensis)的基因组具有全部4类POU,而巴布亚硝水母、海月水母(Aurelia aurita)、水螅只有2类POU。在本研究分析的刺胞动物中,安朵仙水母的POU-1,-6亚型的核苷酸多态性最高;安朵仙水母与指形鹿角珊瑚(Acropora digitifera)、星状海葵的POU-2/3/5亚型的序列多样性较其他钵水母更高。另外,3种钵水母与水螅粘附蛋白的比较结果表明,安朵仙水母具有巴布亚硝水母和海蜇所不具有的粘附相关鼠李糖结合凝集素和一类抗氧化活性物质铁螯合物还原酶。综上所述,安朵仙水母具有更多POU编码基因和复杂POU亚型,以及具有粘附相关凝集素和还原酶的编码基因,可能是与安朵仙水母倒立附着生活方式相关的关键遗传信息。  相似文献   

罗非鱼源无乳链球菌(Streptococcus agalactiae)新型AI-2信号分子受体RbsB蛋白结晶生长研究   总被引：1，自引：0，他引：1  

樊博琳  潘丽霞  王忠良  黎源  简纪常  王蓓 《海洋与湖沼》2020,51(1):132-140

为了开展无乳链球菌(Streptococcus agalactiae)核糖结合蛋白(Ribose binding protein B,RbsB)结构功能的研究,本实验根据已知无乳链球菌ZQ0910全基因组序列设计相关引物。采用PCR方法扩增其RbsB基因,随后将该基因定向克隆到原核表达载体pGEX-6p-1中,在大肠杆菌BL21(DE3)感受态细胞中进行IPTG诱导表达;采用HRV 3C蛋白酶切除GST标签,分子筛分离获得RbsB蛋白;运用生物信息学软件对RbsB基因序列进行分析,并对RbsB蛋白二级和三级结构进行预测;采用NeXtal Tubes JCSG Core Suite结晶试剂盒筛选蛋白结晶条件。研究结果表明,该基因全长为969碱基,编码322个氨基酸,RbsB蛋白理论分子量33.9ku,等电点为9.41,二级结构中α螺旋结构所占比重最高,建立RbsB蛋白三维结构模式图;经IPTG诱导后表达的融合蛋白分子量为59ku,筛选RbsB蛋白的初始结晶条件为(0.2mol/L di-Potassium hydrogen phosphate,20%(W/V)PEG3350;1.5mol/L ammonium sulfate,25%(V/V)Glycerol),获得RbsB蛋白结晶体。本研究结果可为无乳链球菌核糖结合蛋白(RbsB)的功能解析提供实验及理论基础。  相似文献   

Seasonal Changes in the Biochemical Composition of the Edible Sea Urchin Paracentrotus lividus Echinodermata: Echinoidea in a Lagoonal Environment   总被引：1，自引：0，他引：1  

C. Fernandez 《Marine Ecology》1998,19(1):1-11

Abstract. Monthly soluble and insoluble protein, carbohydrate, lipid and ash levels and energetic content were measured in tissues of Paracentrotus lividus adults living in a lagoonal environment over a 1-year period to increase our understanding of resource allocation to the different tissues. The biochemical composition of P. lividus individuals in this environment is similar to that observed in other environments. The gonad and gut contained a high level of proteins soluble or insoluble, a lesser amount of lipids and, finally, a low level of carbohydrates. The main component of the test was ash, followed by protein levels. The lipid and carbohydrate levels in the test were very low. Seasonal variability in the biochemical composition of the different body parts was observed. In the gonad, an increase in the level of lipids was recorded concomitant to an increase in gonad weight. The lipids and carbohydrates, after an initial increase, decreased with the maturation of the gonad. The biochemical variations observed in the gut seem to be linked to both the reproductive cycle and the sea urchin's trophic activity. Variability in the test was low and irregular for all of the observed biochemical components. As P. lividus is an important member of the benthic community in the Urbinu lagoon, this echinoid represents an important store of materials and energy within this lagoonal environment.  相似文献   

Variability in the speciation of iron in the northern North Sea     

Martha Gledhill  Constant M. G. van den Berg  Rob F. Nolting  Klaas R. Timmermans 《Marine Chemistry》1998,59(3-4)

Variations in the speciation of iron in the northern North Sea were investigated in an area covering at least two different water masses and an algal bloom, using a combination of techniques. Catalytic cathodic stripping voltammetry was used to measure the concentrations of reactive iron (Fe_R) and total iron (Fe_T) in unfiltered samples, while dissolved iron (Fe_D) was measured by GFAAS after extraction of filtered sea water. Fe_R was defined by the amount of iron that complexed with 20 μM 1-nitroso-2-napthol (NN) at pH 6.9. Fe_T was determined after UV-digestion at pH 2.4. Concentrations of natural organic iron complexing ligands and values for conditional stability constants, were determined in unfiltered samples by titration. Mean concentrations of 1.3 nM for Fe_R, 10.0 nM for Fe_T and 1.7 nM for Fe_D were obtained for the area sampled. Fe_R concentrations increased towards the south of the area investigated, as a result of the increased influence of continental run off. Fe_R concentrations were found to be enhanced below the nutricline (below 40 m) as a result of the remineralisation of organic material. Enhanced levels of Fe_T were observed in some surface samples and in samples collected below 30 m at stations in the south of the area studied, thought to be a result of high concentrations of biogenic particulate material and the resuspended sediments respectively. Fe_D concentrations varied between values similar to those of Fe_T in samples from the north of the area to values similar to those of Fe_R in the south. The bloom was thought to have influenced the distribution of both Fe_R and Fe_T, but less evidence was observed for any influence on Fe_R and Fe_D. The concentration of organic complexing ligands, which could possibly include a contribution from adsorption sites on particulate material, increased slightly in the bloom area and in North Sea waters. Iron was found to be fully (99.9%) complexed by the organic complexing ligands at a pH of 6.9 and largely complexed (82–96%) at pH 8. The ligands were almost saturated with iron suggesting that the ligand concentration could limit the concentration of iron occurring as dissolved species.  相似文献   

Production of methyl bromide and methyl chloride in laboratory cultures of marine phytoplankton II     

M. G. Scarratt  R. M. Moore 《Marine Chemistry》1998,59(3-4)

Methyl halides (monohalomethanes), especially methyl bromide, are known to contribute significantly to ozone destruction in the stratosphere. Budgets of natural and anthropogenic methyl bromide suggest that marine organisms may be the source of a significant proportion of the total global production. Since phytoplankton are abundant in surface waters, they are obvious candidates. Cultures of nine phytoplankton species were grown in CO₂-enriched, nitrate-limited medium in sealed glass vessels. Species tested include Chaetoceros calcitrans, Isochrysis sp., Porphyridium sp., Synechococcus sp., Phaeodactylum tricornutum, Tetraselmis sp., Prorocentrum sp., Emiliania huxleyi and Phaeocystis sp. Methyl bromide (CH₃Br) and methyl chloride (CH₃Cl) concentrations were determined by bubbling the cultures with high-purity air, cryotrapping the effluent and analyzing it on a gas chromatograph with an electron capture detector. The Phaeocystis sp. samples were monitored with GCMS. Cell population, bacterial population, Chl a, pH, and nitrate concentration were monitored for periods of at least two weeks. CH₃Cl was produced by all cultures. CH₃Br was absent in Tetraselmis sp. and Isochrysis sp. cultures, but present in all the others. Methyl iodide (CH₃I) was present in most cultures but could not be quantified due to analytical limitations. CH₃I and CH₃Br production was fastest in stationary phase and continued long after cell division had ceased. Axeic cultures of Phaeocystis sp. achieved almost identical production rates of CH₃Cl and CH₃Br as xenic cultures of the same species. Species from tropical waters had faster CH₃Cl and CH₃Br production rates than temperate species. Scaling the observed production rates using global standing stock estimates for Chl a and particulate nitrogen indicate that phytoplankton can account for only a fraction of the CH₃Cl and CH₃Br believed to be produced in the ocean. Calculations based on the estimated global biomass of Phaeocystis sp. and E. huxleyi blooms show that their contribution is insignificantly small.  相似文献   

Induction of heat shock protein (hsp)60 in Isochrysis galbana exposed to sublethal preparations of dispersant and Prudhoe Bay crude oil     

M. F. Wolfe  H. E. Olsen  K. A. Gasuad  R. S. Tjeerdema  M. L. Sowby 《Marine environmental research》1999,47(5):517

Adaptation to sublethal exposure to crude oil by phytoplankton is poorly understood. Use of chemical dispersants for oil spill remediation increases petroleum hydrocarbon concentrations in water, while exposing marine organisms to potentially toxic concentrations of dispersant. Heat shock proteins (hsps) have been found to serve as an adaptive and protective mechanism against environmental stresses. The objective of this project was to examine the induction of hsps in Isochrysis galbana, a golden-brown algae, following exposure to the water-accommodated fraction (WAF) of Prudhoe Bay crude oil (PBCO) and PBCO chemically dispersed with Corexit 9527® (dispersed oil: DO). Initial experiments using ³⁵S-labeled amino acids and 2-dimensional electrophoresis with subsequent western blotting identified and confirmed hsp60, a member of the chaperonin family of stress proteins, as being efficiently induced by heat shock in this species. One-dimensional SDS PAGE and western blotting, with hsp60 antibodies and chemiluminesence detection, were used to quantitate hsp60 following exposure to a range of environmental temperatures and concentrations of WAF and DO preparations. I. galbana cultured in 22 parts per thousand (‰) salinity showed a statistically significant increase (p<0.05) in hsp60 after exposure to 25, 30, and 35°C for 1 h compared to controls (20°C), while 34‰ cultures showed no significant increase in hsp60 with increasing temperature. Cultures (22 and 34‰) exposed to varying doses of WAF and DO exhibited statistically significant (p<0.05) increases in levels of hsp60 in all cases. A dose-related response was observed in all exposures (r²≥0.90) with the exception of 34‰ WAF. In independent exposures, a dose-related hsp60 response to naphthalene, a relatively abundant water-soluble aromatic hydrocarbon, was observed in both 22 and 34‰ cultures. Results of this study are consistent with previous studies in other species documenting increases in hsp60 levels with exposure to xenobiotics. Further studies are investigating the protective function of hsp60 against the toxic effects of exposure to WAF and DO preparations.  相似文献   

以哈维氏弧菌(Vibrio harveyi)为重点的鱼类细菌病原的诊断与控制(英文)   总被引：2，自引：0，他引：2  

Ｂ．Ａｕｓｔｉｎ  Ｄ．Ａ．Ａｕｓｔｉｎ  Ｚ．Ｆａｒｈａｔ  Ｐ．Ａ．Ｗ．Ｒｏｂｅｒｔｓｏｎ 《中国海洋大学学报(自然科学版)》1999,(3)

作者对鱼类细菌病原的诊断与控制作了较全面的综述。细菌病原诊断技术包括酶联免疫吸附技术（ＥＬＩＳＡ）及蛋白印迹法（Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ）。细菌病原控制技术包括化学疗剂的应用,有益微生物的应用,营养添加剂,β－葡萄糖苷增强免疫抗病力,及免疫保护作用等。  相似文献   

东风螺、弯竹蛏和斑节对虾的壳基质蛋白组学分析          下载免费PDF全文

张爱华  卫子嫣  姚宇阗  张道锋  黄建科  刘闯 《海岸工程》2022,41(3):258-267

从软体动物中的腹足纲、双壳纲以及节肢动物中的甲壳纲分别选取了3种中国沿海常见的品种——东风螺、弯竹蛏、斑节对虾,提取并分析它们的壳基质蛋白。利用液相色谱串联质谱（LC-MS/MS）技术对蛋白质组学进行分析发现特定的蛋白质具有明显的种属差异：①N66蛋白（一种碳酸酐酶）通常存在于弯竹蛏中,但不存在于东风螺和斑节对虾中,揭示N66蛋白质具有特异性,在未来的物种鉴定中可能具有重要的参考意义;②角质层蛋白AM1199和CP14.6存在于斑节对虾的表皮结构中,在东风螺和弯竹蛏中未检测到;③血蓝蛋白同时在3种动物外壳的基质蛋白中被鉴定出来,揭示了其在不同物种的外壳形成与发育过程中的重要作用。该研究为壳基质蛋白在软体动物或节肢动物外壳中的鉴定提供了基础,也为探索不同物种的生物矿化过程提供了重要参考。  相似文献   

泥蚶(Tegillarca granosa)重组铁蛋白富集重金属离子的特性及化学传感器的研究     

张涛  苏倡  刘艳  张迪骏  周君  芦晨阳  明庭红  司开学  苏秀榕 《海洋与湖沼》2017,48(4):870-876

利用泥蚶(Tegillarca granosa)铁蛋白原核表达工程菌获得的重组铁蛋白,通过圆二色光谱分析蛋白二级结构,扫描电镜和电感耦合等离子质谱研究重组铁蛋白富集Fe~(2+)、Mn~(2+)、Cd~(2+)、Cr~(3+)、Hg~(2+)、Pb~(2+)和As~(3+)等7种重金属离子的特性,同时探索利用重组铁蛋白修饰丝网印刷电极,设计和制备重组铁蛋白检测Pb~(2+)和Cd~(2+)浓度的电化学生物传感器。结果表明复性成功的铁蛋白多为完整的?-螺旋结构,而复性不成功的蛋白聚集体则多为无规卷曲。重组铁蛋白的直径和形态与富集的离子种类有关。泥蚶铁蛋白对单一金属离子Fe~(2+)和Mn~(2+)的富集凸显优势。对两种混合金属离子的富集大多表现为竞争关系。但重组铁蛋白对Hg~(2+)和As~(3+)混合组的富集量明显高于单一金属离子组的富集量,对Hg~(2+)和As~(3+)混合组的富集表现出协同促进作用。重组铁蛋白传感器对Pb~(2+)和Cd~(2+)溶液的最低检测限为10μg/L。  相似文献