海洋微藻胞外产物研究进展

厦门大学国家教育部海洋生态环境开放研究实验室(MEE)开放基金(MEE9903);; 教育部留学回国人员科研启动基金(教外司留[1999])363号资助项

口蹄疫病毒3AB基因的表达及活性分析

通过重叠PCR合成口蹄疫病毒3AB基因,构建原核表达载体pGEX-5X-1/3AB,转化大肠杆菌BL21(DE3),IPTG诱导表达。重组蛋白主要以包涵体的形式表达,将包涵体洗涤溶解后,采用Na2+离子金属螯合亲和层析柱纯化,逐步透析法复性。ELISA实验表明,目的蛋白能与猪口蹄疫阳性血清发生特异性反应。本研究为建立以基因工程产品为抗原、鉴别诊断自然感染和免疫动物的方法提供了技术条件

Spectroscopic characterization and properties of some bioactive peroxovanadium complexes in aqueous solution

Four bioactive peroxovanadium (pV) complexes-bpV(ox), bpV(bipy), bpV(phen). and bpV(pic), ([VO(O-2)(2)L](n-), where ligand L = oxalic acid dianion (ox), bipyridine(bipy), 1,10-phenanthroline(phen), and pyridine-2-carboxylic acid (pic), were synthesized,and characterized by V-51 NMR, H-1 NMR, C-13 NMR, ESI-MS, IR and elemental analysis. All H-1 and C-13 peaks were,assigned by 2D H-1-H-1 peaks were assigned by 2D H-1-H-1 COSY, HMQC and HMBC. Their stereochemical structures in solution were discussed according to the NMR signals of organic ligands. The descending stability order of complexes in aqueous solution determined by V-51 NMR is bpV(phen), bpV(bipy) bpV (pic) and pV(ox). The predominant decomposition patterns of these complexes were proposed on the basis of electrospray ionization MS (ESI-MS) and V-51 NMR. This work will facilitate the studies of interactions between pV complexes and target biomolecules in solution so as:to clarify structure-function relationship of these:bioactive complexes

一种定量检测人血清高敏C反应蛋白的化学发光免疫方法

旨在建立一种可定量检测人血清高敏CRP的化学发光检测方法(High-sensitivity C-reactive protein quantifiable chemiluminescent immunoassay,hs-CRP CLIA)。首先利用亲和层析和离子交换层析技术从肝硬化病人腹水中纯化出高纯度的天然CRP作为免疫原制备了22株CRP单克隆抗体(单抗),其中13株单抗在磷酸胆碱配体捕获ELISA中呈阳性,然后利用方正滴定法筛选出单抗10C5和10C11建立了hs-CRP CLIA。试剂盒评估结果显示:该方法对血清中干扰物质IgG、血红蛋白、甘油三酯等无非特异性反应;该方法检测灵敏度高,在0.04~20.38mg/L范围内定量检测人血清CRP标准品呈良好线性关系(R2>0.993);该方法准确性高、可重复性好,平均回收率为99%,批内差为4.2%~5.8%,批间差为9.0%~11.5%;该方法与进口商品化高敏CRP ELISA试剂盒平行比较检测90份血清标本,结果显示两者有良好的可比性(r=0.968)。综上,建立的hs-CRP CLIA是一种准确、可靠、可定量的高灵敏C反应蛋白检测方法,该方法的临床应用,有利于改善我国心脏病风险评估及肠炎性疾病预后判断

Characterization and immunoprotective effect of SjIrV1,a 66 kDa calcium-binding protein from Schistosoma japonicum

钙结合蛋白是日本血吸虫生长发育不可缺少的蛋白,具有非常广泛而重要的功能。在课题组日本血吸虫体被表膜蛋白研究基础上,利用PCr技术克隆了中国大陆株日本血吸虫66 kdA钙结合蛋白(SJIrV1)编码基因的CdnA序列,blAST分析与菲律宾株日本血吸虫SJIrV1 CdnA编码序列一致,荧光定量PCr分析表明该基因在童虫和成虫期不同发育阶段均有表达,其中在35 d和42 d成虫中表达量较高,在42 d雌虫中该基因表达水平远高于42 d雄虫。构建重组表达质粒PET28A(+)-SJIrV1,在大肠杆菌中成功诱导表达,重组蛋白主要以可溶性形式存在,通过高效液相色谱法(rP-HPlC)以及串联质谱法(MS/MS)鉴定所获蛋白为目的蛋白SJIrV1。蛋白质印迹(WESTErn blOTTIng)分析结果显示重组蛋白能被感染日本血吸虫鼠血清和免疫鼠血清所识别,SJIrV1蛋白在虫体各发育阶段中均表达。免疫荧光染色实验观察表明SJIrV1主要分布在日本血吸虫成虫的表膜。应用重组蛋白免疫bAlb/C小鼠后,免疫鼠血清中检测到较高水平的特异性Igg、Igg1和Igg2A抗体。结果表明SJIrV1可能在日本血吸虫的生长发育过程中起着重要作用。Calcium-binding protein is an indispensable protein which performs extensive and important functions in the growth of Schistosoma japonicum.Based on our primary study on tegument surface proteins of S.japonicun,a cDNA encoding a 66 kDa calcium-binding protein of S.japonicum(Chinese strain) was cloned,sequence analysis revealed that it was identical with that of SjIrV1 of Philippines strains S.japonicum.The expression of SjIrV1 were detected by Real-time PCR,using cDNA templates isolated from 7,14,21,28,35 and 42 days worms and the results revealed that the gene was expressed in all investigated stages,and the mRNA level of SjIrV1 is much higher in 42 d female worms than that in 42 d male worms.The cDNA containing the open reading frame of IrV1 was subcloned into a pET28a(+) vector and transformed into competent Escherichia coli BL21 for expression.The recombinant protein was purified using a Ni-NTA purification system,and confirmed by high performance liquid chromatography(RP-HPLC) and tandem mass spectrometry(MS/MS).Western blotting analysis showed that recombinant SjIrV1(rSjIrV1) could be recognized by the S.japonicum infected mouse serum and the mouse serum specific to rSjIrV1,respectively.Immunofluorescence observation exhibited that SjIrV1 was mainly distributed on the tegument of the 35-day adult worms.ELISA test revealed that IgG,IgG1 and IgG2a antibodies are significantly increased in the serum of rSjIrV1 vaccinated mice.The study suggested that rSjIrV1 might play an important role in the development of S.japonicum.国家自然科学基金(No.31172315); 上海科技发展基金(No.12140902700); 中国博士后科学基金(No.2012M510630)资助~

the physiobiochemistry and molecular marker of three coccolithophorids

球石藻（Coccolithophorids）是细胞外表面具有球石粒（Coccolith）的一类海洋浮游植物的统称，是大洋和近海生态系统的重要组成部分。本文研究了不同环境因子对三种球石藻Emilianiahuxleyi(Lohmann)HayandMohler、Coccolithusneohelis(McIntyreandBé)Reinhardt、Coccolithuspelagicus(Wallich)Schiller的生长特性、生化组成，胞外产物（EPS）的影响，初步观察了Emilianiahuxleyi的细胞形态并研究了Emilianiahuxleyi与其它生物的关系。主要结果如下： 1...Coccolithophorids, which are a group of marine phytoplanktons with coccolith in their extracellular surface, are the one of important components of the ocean and coastal ecosystem. In this present study, the effect of different environmental factors on the growth characteristics、biochemical compostion、EPS of three coccolithophorids (Emiliania huxleyi(Lohmann) Hay and Mohler、Coccolithus neohelis (M...学位：理学硕士院系专业：生命科学学院生物学系_植物学学号：20002600

2-取代-3-烷基异吲哚-1-酮的不对称合成

国家自然科学基金（29832020

Target tracking by initial template and update template

目标跟踪是计算机视觉中的一个重要课题。文中提出一种方法,用于解决跟踪过程中由于目标的运动、变形和外貌变化所引起的跟踪困难问题。该方法基于灰度差平方和,通过优化迭代,求解出当前图像中目标的位置,且考虑利用了目标的先验知识,用初始模板和更新模板同时跟踪,再通过选择误差小的结果作为最终的跟踪目标。仿真实验的结果表明,该方法对于目标的运动、变形和外貌变化以及遮挡,能更稳定地实现跟踪