姜黄素对酵母细胞影响的荧光分析

以最简单的真核生物——酿酒酵母为材料,利用姜黄素的荧光性质,借助荧光光谱法和荧光显微镜研究了姜黄素在酵母细胞中的定位及其对酵母生长的影响.结果表明,姜黄素可以进入酵母细胞中并抑制酵母生长,初步得到了姜黄素对酵母作用的量效关系和时间效应结果,证明了姜黄素可以与酵母细胞线粒体结合.本文结果为以酵母为模型研究姜黄素通过线粒体影响细胞代谢及其与细胞凋亡的关系奠定了基础.国家自然科学基金(批准号:81373296);;厦门大学细胞应激生物学国家重点实验室开放课题(批准号:SKLCSB2017KF003)资助~

Preparation and identification of polyclonal antibody raised against heat shock protein 90 of fission yeast

作者简介:李文珠(1983－),女,博士研究生,主要研究方向:细胞生物学 通信联系人：靳全文，教授，主要研究方向：细胞周期调控及表观遗传学， [email protected][中文文摘]为制备兔抗裂殖酵母Hsp90多克隆抗体,在通过PCR获得了裂殖酵母Hsp90(Swo1)基因后,构建了pMALc2x-Swo1表达载体,可用于表达编码正确氨基酸序列的目的基因。转化大肠杆菌BL21(DE3),IPTG诱导表达,Amylose Resin柱纯化。目的蛋白表达量占菌体总蛋白的30%以上。纯化后,蛋白纯度达95%以上。纯化后的MBP-Swo1融合蛋白抗原加福氏完全佐剂背部皮内注射首次免疫新西兰大白兔,第28天用MBP-Swo1融合抗原加福氏不完全佐剂同样剂量加强免疫,第35天时再次免疫。第49天心脏采血。收集血清后,用免疫印迹(westernblot)检测Swo1多克隆抗体的特异性。免疫印迹检测结果显示该抗体能够特异性识别内源性的裂殖酵母Hsp90/Swo1蛋白,但并不识别人类细胞中的Hsp90α/β。[英文文摘]To prepare the polyclonal antibody of Hsp90(Swo1) of fission yeast Schizosaccharomyces pombe,the encoding gene swo1+was amplified from the yeast genome and then inserted into expression vector pMALc2x.The resulted plasmid pMALc2x-Swo1 was transformed and expressed in E.coli BL21(DE3).The expressed fusion protein was purified through Amylose Resin column.The proteins were expressed mainly as secretion with the yield of more than 30% of total bacterial proteins.After purification,the purity of the proteins was about 95%. The New Zealand white rabbits were immunized with Freund’s complete adjuvant plus purified MBP-Swo1 fusion antigen through the back skin intradermal injection for the first time. The same dose of Freund’s incomplete adjuvant plus MBP-Swo1 was injected to strengthen the immunity after 28 and the 35 days respectively. After 49 days, blood sample was collected from heart, and antisera were extracted. The specificity of Swo1 polyclonal antibody was examined by Western blot. It showed that the antibody obtained had a high specificity to detect endogeneous Swo1 from fission yeast, but it could not recognize Hsp90α/β in human cells. Key words: fission yeast；Hsp90/Swo1；polyclonal antibody.高等学校博士学科点专项科研基金资助项目(20100121110003);国家自然科学基金资助项目(30871376);教育部科学技术研究重点项目(108076

Theoretical Study on the Electronic Spectra of cis-HOOOH and trans-HOOOH

用密度泛函方法(DFT)和全活化空间自洽场方法(CASSCF)以及耦合簇理论(CCSD)优化了反式和顺式HOOOH的平衡几何构型,用DFT计算了HOOOH顺反异构化反应的势能曲线和谐振动频率.用含时密度泛函理论(TD-DFT)和二阶全活化空间微扰理论(CASPT2)计算了反式和顺式HOOOH垂直激发能.计算结果表明:(1)反式异构体比顺式异构体稳定;(2)两种稳定构型的异构化反应有两种路径;(3)对于垂直跃迁能最低的单态和叁态,反式的垂直跃迁能比顺式的低;(4)在单激发态中,CASPT2方法预测的顺式HOOOH寿命最长的激发态为21A″,其跃迁能是167.43nm,寿命为1.44×10-5s;反式HOOOH寿命最长的激发态为21A,其跃迁能是165.52nm,寿命为2.07×10-5s.Equilibrium geometries of cis-HOOOH and trans-HOOOH have been investigated using the density functional theory (DFT), complete active space self-consistent-field (CASSCF), and coupled cluster with single and double replacement (CCSD) approaches. The harmonic vibrational frequencies on the optimized geometries were calculated using the DFT theory. The potential energy curve of the isomerization between the trans-HOOOH and cis-HOOOH was obtained by DFT calculations. Time-dependent density functional theory (TD-DFT) and complete active space perturbation theory of second order (CASPT2) calculations have been performed to obtain the vertical excitation energies of selected low-lying singlet and triplet excited states. Computed results show that: (1) trans-isomer is more stable than cis-isomer; (2) there are two pathways of the conversion between the trans-HOOOH and cis-HOOOH; (3) the vertical excitation energies of the lowest singlet and triplet excited states in trans-HOOOH are lower than those in cis-isomer; (4) in the singlet excited states, 21A state in trans-HOOOH and 21A″ state in cis-HOOOH have the longest lifetimes of 2.07×10-5 s and 1.44×10-5 s with the excitation energies of 165.52 and 167.43 nm, respectively.国家自然科学基金(20473062,20233020,20021002,20173042);; 厦门大学固体表面物理化学国家重点实验室开放课题基金(200306);; 河南省自然科学基金(0311011200,200510475012)资助项

DNA barcoding of hydromedusae in northern Beibu Gulf for species identification

水螅水母类是浮游动物群落的重要组成部分,在近岸海洋生态系统物质循环和能量流动中扮演着重要角色。水螅水母类形态结构简单,但其物种的准确鉴定一直是分类工作中的难点。dnA条形码极大地促进了水螅水母物种的快速、准确鉴定。本研究扩增了北部湾北部28种水螅水母的线粒体COI和16S序列,分别为92条和116条;比较了2个基因片段的种内、种间k2P(kIMurA 2-PArAMETEr)遗传距离;构建了基于这2个基因片段的系统发育邻接树(nEIgHbOr-JOInIng PHylOgEnETIC TrEE);并结合矢量分析构建了klEE-dIAgrAM图。结果显示:COI序列的种内遗传距离为0.008±0.005(0–0.033),种间遗传距离为0.298±0.128(0.092–0.597);16S序列的种内遗传距离为0.006±0.010(0–0.047),种间遗传距离为0.394±0.195(0.068–0.898)。2个基因序列在所调查种类中,种内遗传差异均小于种间遗传差异,存在明显的条形码间隔(bArCOdIng gAP)。基于2个基因片段的nJ树均显示,单种所有个体都位于同一独立分枝。研究结果表明,以COI和16S作为dnA条形码均能对北部湾北部常见水螅水母类进行物种鉴定。Being a major component of coastal zooplankton assemblages, hydromedusae play a key role in material recycling and energy flow of marine ecosystems.Species identification is challenging for this group due to their phonetic simplicity.DNA barcoding provides an efficient method for species identification.In the present study, 92 COI and 116 16 S sequences from 28 hydromedusae species were amplified.A neighborjoining phylogenetic tree was constructed based on Kimura 2-parameter genetic distance and further studied using Klee-diagram vector analysis.Intra-specific K2 P genetic distance averaged 0.008±0.005(0–0.033) for COI, and 0.006±0.010(0–0.047) for 16S; inter-specific K2 P genetic distance averaged 0.298±0.128(0.092– 0.597), and 0.394±0.195(0.068–0.898) for COI and 16 S, respectively.An obvious "barcoding gap" was detected for all species in both markers and all individuals of a species clustered together in both the COI and 16 S trees.Further confirmatory evidence was also provided through indicator vector analysis.Hence, both COI and 16 S appear to be accurate and efficient markers for hydromedusae identification in northern Beibu Gulf.国家自然科学基金青年科学基金(41006078); 海洋公益性行业专项(201005012-3;201005015-5); 厦门大学中央高校基本科研业务费专项资金(2010121037); 厦门大学海洋科学基地科研训练及科研能力提高项目(J1210050

Theoretical Study on Structures and Electronic Spectra of Linear Chain Cluster BC_(2n)B (n=1～12)

应用密度泛函理论,在B3LYP/6-31G*水平上优化得到了线性簇合物BC2B(n=1～12,D∞h)的平衡几何构型,并n计算了它们的谐振动频率.在优化平衡几何构型下,通过TD-B3LYP/cc-pvDZ和TD-B3LYP/cc-pvTZ计算,分别得到了n=1～12和n=1～7的X1Σg→11Σ+u电子跃迁的垂直激发能和对应的振子强度.在B3LYP/6-311+G*水平上计算得到+了簇合物BC2B(n=1～12,D∞h)的电离能.基于计算结果,导出了BC2B体系X1Σg→11Σ+u电子跃迁能以及第一电离能+nn与体系大小n的解析表达式.Using density functional theory, the geometries and the vibrational frequencies of linear chain BC2 B (n=1～12, D∞h) have been investigated at the B3LYP/6-31G* level. Time-dependent density func- n tional theory (TD-DFT) has been used to calculate the vertical transition energies and oscillator strengths for X1Σg →11Σ+u transitions of BC2 B with the cc-pvTZ and cc-pvDZ basis sets. At the B3LYP/6-311+G* + n level, the single-point energies of the clusters have been calculated in order to determine the first ionization energies. On the basis of present calculations, the explicit expressions for the size dependence of the excita- tion energy, the first adiabatic ionization energies (AIE) and vertical ionization energies (VIE) in linear car- bon chains were suggested.厦门大学固体表面物理化学国家重点实验室开放课题基金;; 国家自然科学基金;; 河南省自然科学基金(Nos. 20173042; 20233020; 20021002;0311011200)资助项目

Structure and Electronic Spectrum of Linear Carbon Chains LiC_(2n)Li Studied with Density Functional Theory

应用密度泛函理论,在B3LYP/6 31G(d)水平上优化得到了线型簇合物LiC2nLi(n=1 ~10,D∞h)的基态平衡几何构型,并计算了它们的谐振动频率.利用含时密度泛函理论,计算了簇合物LiC2nLi的X1∑ +g→11∑ +u跃迁的垂直激发能,以及相应的振子强度.基于计算结果,建立了跃迁能和体系大小n的解析关系式. 同时也计算了体系的第一绝热电离能,讨论了体系的电离能与体系大小n的关系.The geometries and the vibrational frequencies of linear chains LiC_(2n)Li(n=1~10) were investigated by density functional theory at the B3LYP/631G(d) level. Time-dependent density functional theory was used to calculate the vertical transition energies and oscillator strengths of X~1∑~+_g→1~1∑~+_u transitions in LiC_(2n)Li. Based on the present calculations, the explicit analytic expression between the vertical transition energies and n was obtained. Meanwhile, the first adiabaticionization energies werecalculated, and the relationship between the adiabaticionization energiesand nhas also been discussed.固体表面物理化学国家重点实验室(厦门大学)开放课题基金 (200306);; 国家自然科学基金 (20173042);; 河南省自然科学基金 资助项目(0311011200)

Structure and Electronic Spectrum of Linear Carbon Chain PC_(2n)P Studied with DFT

应用密度泛函理论,在B3LYP/6-31G**和B3LYP/6-311G**水平上优化得到了线型簇合物PC2nP(n=1-10)的基态平衡几何构型,计算了它们的谐振动频率.在基态平衡构型下,利用含时密度泛函理论,计算得到了簇合物PC2nP(n=1-10)的垂直激发能和相应的振子强度,导出了激发能与体系大小n的解析关系式.The geometries and the vibrational frequencies of linear carbon chain PC_ 2nP (n=1-10) were investigated by density functional theory(DFT) at the B3LYP/6-31G ** and B3LYP/6-311G ** levels. Time-dependent density functional theory (TD-DFT) was employed to calculate the vertical transition energies and oscillator strengths. On the basis of present calculations, the explicit expressions for the size dependence of the excitation energy in linear carbon chain was suggested.ThisresearchwassupportedbyStateKeyLaboratoryforPhysicalChemistryofSolidSurfaces(XiamenUniversity)(200306),andNaturalScienceFoundationofChina(20173042,20233020)

An ecological study on zooplankton in the northern Beibu Gulf V: the effects of microzooplankton grazing on phytoplankton in summer

2011年8月份于北部湾北部海域5个观测站位获得的分层水样,分析了表层叶绿素A含量和表层微型浮游动物丰度以及类群组成;同时于现场采用稀释培养法研究了该海域浮游植物生长率(μ)和微型浮游动物的摄食率(g)。分析和测定结果表明:调查海区的微型浮游动物丰度400—1167个/l,类群组成以无壳纤毛虫为主;浮游植物的生长率为-1.50—1.13 d-1,微型浮游动物摄食率为0.33—1.08 d-1;推算微型浮游动物对浮游植物现存量以及初级生产力的摄食压力分别为28.1%—66.0%和-7.4%—438.4%。相对于中国其他海区,8月份北部湾北部海域微型浮游动物摄食速率处于中等水平。调查期间,广西沿海高生产力海区,浮游植物生长率大于微型浮游动物动物的摄食率,浮游植物生物量处于积累期;涠洲岛以南海域,浮游植物生产力较低,微型浮游动物摄食作用是控制浮游植物生长的重要因素。In August 2011,we quantified the impact of microzooplankton grazing on phytoplankton production in the surface waters of the northern Beibu Gulf,Vietnam.Shipboard dilution incubation experiments were carried out at five stations to calculate phytoplankton specific growth rates and the specific rates of grazing losses to microzooplankton.Dilution experiments using chlorophyll a( Chl a) as a tracer were used to estimate daily rates in two size fractions; image-analyzed microscopy provided quantitative estimates of microzooplankton standing stock( including ciliate and vertebrate larvae).PFW( particle-free water) was used to dilute seawater to five target dilutions of 0%,20%,40%,60%,and 80%.Microzooplankton grazing and phytoplankton growth rates were estimated by the linear regression of AGR( apparent growth rate) versus dilution factor.We estimated the grazing impact of microzooplankton on phytoplankton by calculating the percentage of phytoplankton standing stock and potential primary production ingested.Average total chlorophyll a concentration was( 0.67 ± 0.58) μg / L.Small-celled phytoplankton dominated this particular community.A notable characteristic of the phytoplankton community structure was that nano- and picoplankton made up a significant portion.Aloricate ciliates dominated the microzooplankton samples,in particular Strombidium,which accounted for 27.2% of total aloricate ciliate abundance.In addition,loricate ciliates and copepod nauplii were recorded.Microzooplankton densityvaried from 400 to 1167 ind / L.The horizontal distribution of microzooplankton was greater in nearshore than in offshore waters.Abundance did not differ significantly with chlorophyll a concentration in this study; however,there was a positive correlation between microzooplankton abundance and temperature.The phytoplankton growth rate( < 200 μm) ranged between-1.50 and 1.13 d-1,with the highest values being associated with nearshore waters.Meanwhile phytoplankton mortality due to microzooplankton grazing( 0.33—1.08 d-1) was highest at the same nearshore sites; however,growth rates were low at these sites.Consequently,microzooplankton consumed 28.1%—66.0% of chlorophyll a standing stocks and-8.1%—438.4% of the primary production per day.However,the grazing rate of microzooplankton on nanoplankton( 2— 20 μm) varied from 1.87 to 0.52 d-1,with an average of( 0.94±0.57) d-1.Given that the rates of grazing on phytoplankton of <200 μm were lower than those on nanoplankton,we conclude that microzooplankton prefer to graze on nanoplankton.There was a positive correlation between phytoplankton growth rate and grazing rate; in addition,significant differences between grazing rate and chlorophyll a were found.This suggests that microzooplankton grazing activity is the main factor regulating the community structure and restraining the growth of phytoplankton.During the investigation period, microzooplankton grazing effectively controlled the growth of phytoplankton,consuming 176.2% of primary production on average.When grazing rates are far greater than phytoplankton growth rates,energy is transferred from lower to higher trophic levels more effectively.In the surface waters of the northern Beibu Gulf,microzooplankton largely control the growth of phytoplankton by grazing offshore rather than in nearshore waters where primary production is high.The growth and grazing rates combined promote planktonic ecosystem stability in this area.These results indicate that grazing by microzooplankton is a key process controlling the growth of phytoplankton in this area.海湾公益项目(201005012

The relationship between nutrients and phytoplankton community structure in northern Beibu Gulf

根据2011年4月和8月北部湾北部(北部湾20°n以北的海域)的调查数据,北部湾北部春夏两季共鉴定出浮游植物312种,隶属于4门78属,硅藻是主要优势类群。春季优势种为细弱海链藻(THAlASSIOSIrA SubTIlIS)、丹麦细柱藻(lEPTOCylIndruS dAnICuS)、奇异棍形藻(bACIllArIA PArAdOXA)和尖刺拟菱形藻(PSEudOnITzSCHIA PungEnS);夏季优势种仅为海链藻(THAlASSIOSIrA SP.)。浮游植物群落在春季可划分为2个子群落(SPr1和SPr2),在夏季可划分为3个子群落(SuM1、SuM2和SuM3)。该海域n/P比值一直处于高的水平,绝大部分的水域表现为磷限制,这可能是硅藻大量生长所造成的。春季SPr2群落优势种的柔弱角毛藻(CHAETOCErOS dEbIlIS)与除铵盐之外的营养盐具显著相关关系,奇异棍形藻与环境因子无相关关系。夏季SuM3群落优势种的奇异棍形藻与磷酸盐具显著相关关系。分析表明,琼州海峡过道水为北部湾北部输送了磷酸盐,而硅酸盐则主要由陆地径流输入。Phytoplankton community and nutrient concentration in the northern Beibu Gulf in April and August 2011 were investigated.In this research, 312 phytoplankton taxa were identified, which belonged to four phyla and 78 genera, with Bacillariophyta being the main group.The dominant species were Thalassiosira subtilis, Leptocylindrus danicus, Bacillaria paradoxa, and Pseudonitzschia pungens in spring, and Thalassiosira sp.in summer.Phytoplankton community in northern Beibu Gulf could be divided into two natural groups in spring(spr1 & spr2) and three in summer(sum1, sum2 & sum3).N/P ratio was in a relatively high level in the study area, and most of the waters were phosphorus limitated which resulted from diatom blossom.Chaetoceros debilis was the dominant species of group spr1 and had significant correlation with nutrient in spring, except for ammonium.Bacillaria paradoxa was the dominant species of group sum3 and had significant correlation with phosphate in summer, but, it was uncorrelated with any environmental factors as the dominant species of group spr2 in spring.Phosphate was transported to northern Beibu Gulf through the Qiongzhou Strait corridor water, and silicate was input by land runoff.海洋公益性行业科研专项项目(201005012

Ecosystem structure and function in northern Beibu Gulf: zooplankton spatial niche and its differentiation

根据2006—2007年北部湾北部海域4个航次调查资料,采用k-优势度曲线法对浮游动物种(类)群进行排序分析,运用lEVInS公式和PIAnkA指数分析了主要优势种(类)群的生态位宽度值和生态位重叠程度,并通过除趋势典范对应分析(dCCA)研究生态位分化状况。结果表明,该海区浮游动物优势种(类)群按生态位宽度值可划分为广生态位、中生态位和窄生态位三大类型。其中,广生态位的种(类)群如肥胖软箭虫、亚强次真哲水蚤、长尾类幼体等具有较强的生态适应性,生活范围较广;窄生态位的种(类)群,对环境的适应性较差,对海流和水团等环境因素的变化具有指示意义。生态位宽度能够反映种(类)群丰度季节变化,但不能反映具体丰度大小。北部湾北部全年主要优势种(类)群的生态位宽度值介于0.08—0.77之间,并且各优势种(类)群的生态位宽度差异较大,种对间生态位重叠值介于0.02—0.89之间,平均生态位重叠指数为0.45,种(类)群之间利用资源环境的互补性较强,这与海域内生境多样化并且各优势种对环境适应能力的差异存在一定联系。dCCA分析表明,影响浮游动物分布的主要因子是水深、温度和盐度,其次是叶绿素A和溶解氧。The concept of ecological niche had remained as one of the core ideas in ecological research for almost a century,which was closely linked with interspecific competition,resource utilization had played an important role in the study of community composition and function,of the relationships among species,biodiversity,community succession and population evolution.Based on four seasonal investigations( Spring 2007,Summer 2006,Autumn 2007 and Winter 2006) in northern Beibu Gulf from 2006 to 2007,the niche breadth,niche overlap,spatial differentiation and environment effects on the abundance and spatial distribution of zooplankton were studied with qualitative and quantitative methods.The cumulative top 80% of zooplankton dominant populations were selected by the K-dominant curve method,then the niche breadths and their overlaps of zooplankton were measured using the formulas proposed by Levins Niche Breadth index and the Pianka Niche Overlap index,and the spatial niche differentiation was studied by using Detrended Canonical Correspondence Analysis( DCCA) method.The results showed that the zooplankton dominant species in northern Beibu Gulf were classified in to three groups,i e.,wide niche breadth groups( Bi= 0.60—0.77),middle niche breadth groups( Bi= 0.30—0.60),narrow niche breadth groups( Bi= 0.08—0.30) based on the niche breadth value.The niche breadths of zooplankton such as Sagitta enflata,Subeucalanus subcrassus and Macrura larvae were wider,which was related with their own characteristics and the capability of adapting environment,while the species whose niche breadths were narrower such as Doliolum denticulatum and Dolioletta gegenbauri will be considered as an indicative significance for ocean currents and water masses.Niche breadth could reflect the zooplankton abundance with the seasonal dynamic changes,and also could objectively reflect the species in the habitat in which the distributions of degree but it could not reflect the Accurate zooplankton abundance.The niche breadth values of dominance zooplankton species ranged between 0.08 and 0.77 in the whole year and The niche breadth difference among the dominant populations was great; The niche overlap values of dominance zooplankton species ranged between 0.02 and 0.89 in the whole year and the average niche overlap index was 0.45,there was strong complementarity in terms of the utility of resource among zooplankton populations,which had certain relationship with northern Beibu Gulf habitat diversification and the strong complementarity in terms of the utility of resource among zooplankton populations.The Detrended Canonical Correspondence Analysis result showed that depth,temperature and salinity had more important influence on the species spatial distribution than Chlorophyll a and dissolved oxygen.海湾公益项目(201005012); 我国近海海洋综合调查与评价专项(908-01-ST09