Molecular Responses of Plasma Membrane Ca2+-ATPase of Eichhornia Crassipes under Heavy Metal Stress

凤眼莲[Eichhorniacrassipes（Mart.）Solms]，属于雨久花科（Pontederiaceae），凤眼莲属（EichhorniaKunth），多年生的水生草本植物。繁殖能力很强，致河流、湖泊阻塞，被人们列入恶性入侵杂草之列。但研究表明，凤眼莲能有效地去除污水中氮、磷等营养元素吸收和富集各种重金属和有毒化合物等，是一种监测和净化水体环境污染的良好植物材料。目前环境的日益恶化，重金属污染情况越来越严重。本文从凤眼莲PMCa2+-ATPase的活性及相关基因表达的变化方面来研究凤眼莲对重金属的耐受及响应，为我们以后利用凤眼莲的耐污和净化能力提供科学理论基础。 本文在比较了Ge...Eichhornia crassipes, belonging to Eichhornia Kunth, Pontederiaceae, is a perennial aquatic herb. E. crassipes with strong reproductive ability, which can easily disrupt waterways, is considered one of malign invasive plants. But the research indicate that nutrient element(N, P), heavy metal and toxic compounds can be efficiently removed form sewage by Eichhornia crassipes. The study of tolerance ...学位：理学硕士院系专业：海洋与环境学院环境科学与工程系_环境科学学号：2262007115239

Construction of SSH library from haemocyte of variously colored abalone challenged with bacteria and differential expression analysis of macrophage expressed protein

以雌性杂色鲍为对象,以大肠杆菌、副溶血弧菌、溶壁微球菌、表皮葡萄球菌、金黄色葡萄球菌的混悬液做为攻毒菌,利用抑制性差减杂交(SSH)技术构建细菌攻毒的杂色鲍血淋巴细胞SSHcDNA文库。随机挑取生长菌落110个克隆子,进行菌液PCR鉴定,计算文库重组率为98.18%,文库容量为1.37×106pfu。将重组子测序,经BLAST一致性搜索比对分析,有一重组片段含有穿孔素(Perforin)保守结构域,为巨噬细胞表达蛋白(MEP)类穿孔素部分cDNA序列,片段大小为1551bp,连续编码517个氨基酸残基,申请GenBank登录号为EU272049。经半定量PCR和荧光定量PCR差异显示分析,发现在细菌感染状态下MEP基因在血淋巴细胞中存在明显的上调表达现象。Abalones are considered to be the most precious delicacy from the sea, and become very important commercial seafood in aquaculture worldwide. Variously colored abalone (Haliotis diversicolor Reeve, 1846) has been widely cultured on the southeast coast for more than twenty years. However, abalone culture frequently suffers from bacterial infection and mass mortality of reared abalones causes serious economic losses. Unfortunately, knowledge of the defense mechanism in this animal is still lacking. In this study, using suppression subtractive hybridization (SSH) technology, a forward SSH li-brary was constructed from haemocytes of H. diversicolor, with the content of 1.37×106 pfu and the recombinant rate of98.18%. After the recombinant plasmids were sequenced, partial cDNA of macrophage expressed protein (MEP) was recognized based on BLAST searches in NCBI, with the size of 1 551 bp, and continuously encoding 517 amino acids. Semi-quantitative PCR and quantitative real-time PCR results showed that MEP cDNA was distinctly up-regulated in haemocytes of the bacterial-challenged group compared to the unchallenged group. The gene information obtained from this library will provide new insights into the immune mechanism of H. diversicolor and facilitate future study of target genes involved in the response to invading microorganisms.国家高技术研究发展计划项目(863计划)(编号:2007AA091406)资助~

Prokaryotic Expression and Purification of Ca~(2+)-ATPase from Aquatic Plant Eichhornia crassipes

从水生植物凤眼莲叶片中提取总rnA,经rT-PCr扩增出CA2+-ATPASE基因片段,经限制性内切酶(SMA I,nOT I)酶切后按正确的读码框顺序插入到PgEX-4T-2表达载体上,重组质粒转化大肠杆菌,经菌落PCr和质粒双酶切鉴定、序列测定确认,证实成功地构建了CA2+-ATPASE基因融合表达载体,.转化菌经IPTg诱导表达,获得了大小约48kd的可溶性目的蛋白,与预期相吻合.利用谷胱甘肽琼脂糖凝胶4b(gluTATHIOnE SEPHArOSE 4b)亲和介质对重组蛋白进行纯化,获得了高纯度的目的蛋白.The Ca2+-ATPase gene was cloned fromEichhornia crassipes leaves using the PCR technology.After digested by the enzymes(Sma I,Not I),it was inserted into the plasmid pGEX-4T-2to reconstruct the expression vector.The recombinant protein was induced by IPTG and then purified using Glutathione Sepharose 4B.As a result,a single 48 kDa protein was acquired,which implied the protein was the purified Ca2+-ATPase fusion protein.国家自然科学基金面上项目(30770391

Language cognition and language computation — human and ma chine language understanding

语言理解是认知科学和计算机科学交叉领域共同关心的问题,但两个学科在选择具体研究问题时却十分不同.认知科学领域的研究侧重解析大脑的工作机制,更多地关注于描述大脑对语言的响应,缺乏对大脑语言功能整体化、系统化的研究,而计算机科学家在选择研究问题时重点关注实际应用效能,往往忽略了对语言最本质规律的研究.那么,如何实现两种思路的交叉融合,为智能语言计算模型的构建和语言认知机理的研究带来新的机遇和启发呢?本文首先简要回顾了认知科学和计算机科学在语言理解方向上的研究问题、发展历程和研究方法,重点阐述研究现状和面临的挑战,之后对比认知科学和计算机科学领域对于语言理解问题的主要观点,分析两者之间的异同.最后对现有语言认知和语言计算两个领域的交叉融合方法进行归纳和总结,并对未来发展趋势予以展望.</p

Benzo[a]pyrene modulation of acute immunologic responses in red Sea bream pretreated with lipopolysaccharide

The effects of polycyclic aromatic hydrocarbons (PAHs) have been reported to modulate the immune response in aquatic animals, but the collected information of their effects on fish immunity is so far ambiguous. This study demonstrated that Benzo[a]pyrene (BaP) exposure altered the expression pattern of an antimicrobial peptide hepcidin (PM-hepc) gene and the activities of some immune-associated parameters in the lipopolysaccharide (LPS)-challenged red sea bream (Pagrus major). It was observed that LPS could increase respiratory burst, lysozyme and antibacterial activity in P. major. However when the P. major was exposed to different concentrations of BaP (1, 4, or 8 μg L-1) for 14 days and then challenged with LPS there was no significant change in the lysozyme and antibacterial activity. It was further observed that LPS could induce the PM-hepc mRNA expression at 3, 6, and 12-h post-LPS challenge. However, when P. major was exposed first to BaP for 14 days and then challenged with LPS, the expression of PM-hepc mRNA was delayed in the liver until 24 h and not significantly induced until 48 and 96 h. The mRNA expression pattern was completely different from that only with LPS challenge, showing that BaP exposure changed the PM-hepc mRNA expression pattern of fish with LPS challenge. This study demonstrated that BaP exposure can weaken or inhibit the induction of lysozyme and antibacterial activity in the LPS-challenged P. major; conversely BaP exposure could enhance the mRNA expression of PM-hepc gene, indicating that the effect of BaP has different modulatory mechanism on hepcidin genes and immune-associated parameters. ? 2012 Wiley Periodicals, Inc