Molecular Characterization of a Dehydroascorbate Reductase from Pinus bungeana

Dehydroascorbate reductase （DHAR） plays a critical role in the ascorbate-glutathione recycling reaction for most higher plants. To date, studies on DHAR in higher plants have focused largely on Arabidopsis and agricultural plants, and there is virtually no information on the molecular characteristics of DHAR in gymnosperms. The present study reports the cloning and characteristics of a DHAR （PbDHAR） from a pine, Pinus bungeana Zucc. ex Endl. The PbDHAR gene encodes a protein of 215 amino acid residues with a calculated molecular mass of 24.26 kDa. The predicted 3-D structure of PbDHAR showed a typical glutathione S-transferase fold. Reverse transcripUon-polymerase chain reaction revealed that the PbDHAR was a constitutive expression gene in P. bungeana. The expression level of PbDHAR mRNA in P. bungeana seedlings did not show significant change under high temperature stress. The recombinant PbDHAR was overexpressed in Escherichia coil following purification with affinity chromatography. The recombinant PbDHAR exhibited enzymatic activity （19.84 i.mnol/min per mg） and high affinity （a Krn of 0.08 mM） towards the substrates dehydroascorbate （DHA）. Moreover, the recombinant PbDHAR was a thermostable enzyme, and retained 77% of its initial activity at 55℃. The present study is the first to provide a detailed molecular characterization of the DHAR in P. bungeana.     

Molecular characterization of the duck enteritis virus <Emphasis Type="Italic">UL4</Emphasis> gene

Duck enteritis virus (DEV) is a herpesvirus that causes an acute, contagious and fatal disease. In the present article, the DEV UL4 gene was cloned and sequenced from a vaccine virus. A degenerate oligonucleotide primer for the consensus site of herpesvirus UL3 gene and a specific primer located in UL5 were used in the polymerase chain reaction (PCR) to amplify a DNA product 2 086 bp in size. DNA sequence analysis revealed that a 714 bp open reading frame (ORF) of DEV encoding a 237 amino acid polypeptide is homologous to the family of herpesvirus UL4 proteins and therefore has been characterized as a DEV UL4 gene. Alignment of the DEV UL4 protein sequence with those of other alphaherpesviruses showed that 10 amino acid residues are completely conserved. Phylogenetic tree analysis showed that the seventeen alphaherpesviruses viruses analyzed were classified into four large groups, and the duck enteritis virus branched separately, closely related to the Mardiviruses group comprising Gallid herpesvirus 2 (GaHV-2), Gallid herpesvirus 3 (GaHV-3) and Meleagrid herpesvirus 1 (MeHV-1). The present study showed that the evolutionary relationship of the UL4 protein could be used for classification of alphaherpesviruses.      

壳聚糖Fe（Ⅲ）配合物的合成、表征及抑菌性能

合成了壳聚糖Fe(Ⅲ)配合物,利用IR、UV、和TG-DTA对配合物进行了表征.研究了壳聚糖及壳聚糖铁(Ⅲ)配合物对常见的两种细菌(大肠杆菌、金黄色葡萄球菌)的抑菌性能.结果表明:水溶性的壳聚糖在配合前后,其结构表征发生了一定的变化,并且壳聚糖铁(Ⅲ)配合物的抑菌性能比水溶性壳聚糖本身显著提高(P＜0.05).     

流动电位法对甘氨酸水溶液结晶过程的表征

以流动电位法研究甘氨酸饱和水溶液降温结晶过程。考察了溶液酸度和起始温度对结晶过程中流动电位υ-结晶器温度t曲线的影响。结果表明,甘氨酸饱和溶液在较高起始温度30和35℃下成核能力较差,在较低起始温度20和25℃下成核能力较强。随着甘氨酸饱和溶液酸度的变化,相同起始温度的υ-t曲线表明成核能力有较大差异。溶液酸度为pH=7.0、pH=5.0、pH=4.0的条件下成核作用明显,但低温下成核具有较大的偶然性。等电点附近(pH=6.0)成核能力较差,容易析出不定型固体。运用生长基元理论进行了分析。     

阳离子聚丙烯酰胺的合成及其表征

合成了阳离子功能单体甲基丙烯酰氧乙基二甲基丁基溴化铵(DMB),并将DMB与丙烯酰胺(AM)共聚制备了阳离子聚丙烯酰胺,并用IR,NMR对其结构进行了表征。     

翘嘴鳜NADPH氧化酶2个催化亚基cDNA的克隆和表达特征

吞噬细胞NADPH氧化酶能生成用于清除病原微生物的活性氧(reactive oxygen species, ROS),在机体的防御体系中起着非常重要的作用.本文利用RT-PCR结合RACE-PCR的方法,克隆到翘嘴鳜NADPH氧化酶的催化亚基gp91phox和p22phox的cDNA全长.并研究两者在正常的翘嘴鳜和注射了柱状黄杆菌灭活菌苗（FKG4）的翘嘴鳜组织中的表达模式.结果表明,gp91phox基因cDNA序列全长2 037 nt,开放阅读框长度为1 698 nt,翻译成565个氨基酸;p22phox 基因cDNA序列全长1 296 nt,开放阅读框561 nt,翻译成186个氨基酸.将这2个亚基推导的氨基酸序列与人的对应亚基相比,相似性分别为68.7%和60.8%,且具有相似的结构域和功能域,说明翘嘴鳜与人的NADPH氧化酶具有相似的功能活性.半定量PCR分析显示,在翘嘴鳜血液、脑、心脏、肾、肝、脾、胸腺等11种组织中均能检测到gp91phox和p22phox的基因表达.经FKG4免疫后,gp91phox在翘嘴鳜血液、头肾和脾3种组织中的表达量显著上升,p22phox在头肾和脾2种组织中的表达量显著上升.由此推断,NADPH氧化酶可能参与了机体的抗菌免疫应答.     

白藜芦醇分子印迹聚合物微球的制备及特性评价(英文)

以聚苯乙烯微球为种球,白藜芦醇为模板分子,采用单步溶胀聚合法在N,N-二甲基甲酰胺体系中制备了单分散分子印迹聚合物微球。用扫描电镜对微球的结构和形貌进行了表征,并研究了微球的制备条件和吸附特性。微球的凹陷可有效地增加微球的比表面积和结合位点,从而提高了模板分子的结合速率及微球的印迹容量。     

聚乙二醇丙烯酸酯的合成与表征

以聚乙二醇-400（PEG400）与丙烯酸直接缩合反应,在不加有毒带水剂的条件下合成了丙烯酸聚乙二醇酯（PEGA）。通过正交实验确定酯化反应的最佳条件：丙烯酸／PEG400的摩尔比为2．0：1．0,反应温度是110℃,阻聚剂对苯二酚为0．4％（以醇酸总质量计）,反应时间为6小时,催化剂对甲苯磺酸为0．8％（以醇酸总质量计）,产率为76．7％。产品结构经IR和1HNMR表征,证明是所需的产物。     

中国特有金钱槭属植物物种分化研究

金钱槭属(Dipteronia Oliv.)隶属于槭树科(Aceraceae),是北温带植物区系中古老、残遗的木本属之一,为中国特有属,属下仅包括金钱槭(D.sinensis Oliv.)和云南金钱槭(D.dyerana Henry)2个物种。本实验从形态学、分子遗传学及基因序列分析等方面对金钱槭(D.sinensis Oliv.)和云南金钱槭(D.dyerana Henry)进行对比研究,探讨其物种间分化程度。结果表明:(1)形态学研究显示两物种形态性状存在显著差异,表明其在形态上发生了显著分化;(2)基因组DNA的AFLP标记分析表明,两物种间存在极显著遗传分化(ΦST=0.390 4),在物种水平及群体水平,两物种遗传多样性及遗传变异水平皆有明显差异(金钱槭云南金钱槭);(3)从GenBank数据库中选出2物种已知的叶绿体基因(psbM-trnD、trnD-trnT、trnL、trnL-trnF、rpl16)和核基因ITS(ITS1、5.8SRNA、ITS2)进行序列进化比对分析,t-test检验表明,2物种叶绿体基因序列间有极显著差异。研究结果表明:金钱槭和云南金钱槭两物种间产生了极显著的分化,推测可能由于长期的地理隔离所致。     

Characterization of Leaf-Inspired Microfluidic Chips for Pumpless Fluid Transport

Microfluidic networks are extensively used in miniaturized lab-on-a-chip systems. However, most of the existing micro- channels are simply designed and the corresponding microfluidic systems commonly require external pumps to achieve effec- tive fluid transport. Here we employed microfabrication techniques to replicate naturally-optimized leaf venations into synthetic hydrogels for the fabrication of pumpless microfluidic chips. The unique properties of leaf-inspired microfluidic network in convectively transporting fluid were characterized at different inclination angles. Flow velocity inside these microfluidic net- works was quantitatively measured with Particle Image Velocimetry （PIV）. Mass diffusion from biomimetic microfluidic network to surrounding bulk hydrogels was investigated. The results demonstrate that the leaf-inspired microfluidic network can not only effectively transport fluid without the use of external pumps, but also facilitate rapid mass diffusion within bulk hy- drogel chips. These leaf-inspired microfluidic networks could be potentially used to engineer complex pumpless or- gan-on-a-chip systems.