SARS病毒E基因的克隆及原核表达

为了得到纯的SARS-CoV E蛋白以进一步研究其功能,通过Touch-down PCR程序,从SARS-CoV WHU cDNA(pMD18-T载体)文库中扩增了E基因,构建了重组质粒pGEX-E,并在Ecoli DH5α中进行了原核表达。SDS/PAGE及Western blot结果表明SARS病毒E蛋白的分子量约5 kDa,较预测的略小,并进一步分析了这种现象产生的可能原因。     

基于全蛋白质组的微生物系统发育树构建

新近出现的信息离散性度量方法(简称FDOD方法)已在多个领域获得成功的应用,是一种非比对距离方法.随着越来越多的微生物全基因组测序任务的完成,人们开始在整个基因组水平上探讨物种的系统发育关系.因此,将FDOD方法应用于微生物系统发育分析是一项很有意义的工作.因为氨基酸序列比DNA序列更为保守,能为物种的进化分析提供更为有用的信息.对收集到的163个原核生物和5个真核生物,从完全蛋白质组出发去分析推断其系统的发育关系,所得的系统发育树包括145个细菌、18个古细菌和5个真核细菌,这与三界进化理论符合,大部分低层分支与权威的《伯杰氏系统细菌学手册》相同.并且对高层分支关系提出了一些新建议.     

Identification of encoding proteins related to SARS-CoV

By sampling I00 encoding proteins from SARS-coronavirus (SARS-CoV, NC 004718) and other six coronaviruses and selecting 23 variables through stepwise multiple regression (SMR) from 172 variables, the multiple linear regression (MLR) model was established with good results of the quantitative modelling correlation coefficient R^2=0.645 and the cross-validation correlation coefficient Rcv^2=0.375. After removing 4 outliers, the quantitative modelling and cross-validation correlation coefficients were R^2=0.743 and Rcv^2=0.543, respectively.     

Molecular phylogeny of coronaviruses including human SARS-CoV

The outbreak of SARS sets an urgent task to reveal the origin of human SARS-CoV, i.e. its relation to other known species of coronavirus, and to trace the genetic variation in the spreading process of SARS. Partial answer to the problem may be obtained from phylogenetic analy-sis of available genomes. We call a phylogenetic tree of different species of coronavirus including the human SARS-Cov a CoV Tree and that of different isolates of SARS-CoV a SARS Tree. CoV trees have been c…     

A complete sequence and comparative analysis of a SARS-associated virus(Isolate BJ01)

Severe Acute Respiratory Syndrome (SARS) is a newly identified infectious disease[1—5]. The global outbreak of SARS has been threatening the health of people worldwide and has killed 353 people and infected more than 5462 in 27 countries, as reported by WHO on April 29, 2003 (http://www.who.int/csr/sarscountry/en). Although it has been recognized that a variant of virus from the family of coronavirus might be the candidate pathogen of SARS[1—5], its identity as the unique pathogen sti…     

A complete sequence and comparative analysis of a SARS-associated virus （Isolate BJO1）

The genome sequence of the Severe Acute Respiratory Syndrome (SARS)-assoclated virus provides essential information for the identification of pathogen(s), exploration of etiology and evolution, interpretation of transmission and pathogenesis, development of diagnostics, prevention by future vaccination, and treatment by developing new drugs.We report the complete genome sequence and comparative analysis of an isolate (B J01) of the coronavirus that has been recognized as a pathogen for SARS. The genome is 29725 nt in size and has 11 ORFs (Open Reading Frames). It is composed of a stable region encoding an RNA-dependent RNA polymerase (composed of 20RFs) and a variable region representing 4 CDSs (coding sequences) for viral structural genes (the S, E, M, N proteins) and 5 PUPs (putative uncharacterized proteins). Its gene order is identical to that of other known coronaviruses. The sequence alignment with all known RNA viruses places this virus as a member in the family of Coronaviridae. Thirty putative substitutions have been identified by comparative analysis of the 5 SARS-associated virus genome sequences in GenBank. Fifteen of them lead to possible amino acid changes (non-synonymous mutations) in the proteins. Three amino acid changes, with predicted alteration of physical and chemical features, have been detected in the S protein that is postulated to be involved in the immunoreactions between the virus and its host.Two amino acid changes have been detected in the M protein,which could be related to viral envelope formation. Phylogenetic analysis suggests the possibility of non-human origin of the SARS-associated viruses but provides no evidence that they are man-made. Further efforts should focus on identifying the etiology of the SARS-associated virus and ruling out conclusively the existence of other possible SARS-related pathogen(s).     

SARS相关冠状病毒刺突糖蛋白的研究(综述)

概述了SARS—CoV的S蛋白的结构及功能相关研究。严重急性呼吸综合症相关的冠状病毒(SARS-CoV)引起2003年我国南方非典型肺炎爆发流行,波及多个国家和地区。目前全球许多学对SARS-CoV进行了广泛的研究,发现S蛋白是病毒表面的主要蛋白,它构成冠状病毒科特征性的冠状样结构,在严重急性呼吸综合症的发病机制起着关键性作用,可介导表达相关受体的宿主细胞感染。现已鉴定出SARS-CoV的S蛋白相关受体,同时它在抗病毒感染中是一个关键靶蛋白。     

Phylogenetic positions of two marine ciliates, Metanophrys similis and Pseudocohnilembus hargisi (Protozoa, Ciliophora,Scuticociliatia), inferred from complete small subunit rRNA gene sequences

The small subunit rRNA (SSrRNA) gene was sequenced for two marine scuticociliates Metanophrys similis and Pseudocohnilembus hargisi. The results show that this gene comprises 1763 and 1753 nucleotides in the two marine ciliates respectively. Metanophrys similis is phylogenetically closely related to the clade containing Mesanophrys carcini and Anophyroides haemophila, which branches basally to other species within the order Philasterida. Pseudocohnilembus hargisi groups with its congener, P. marinus, with strong bootstrap support. Paranophrys magna groups with the clade including Cohnilembus and Uronema, representing a sister clade to that containing the two Pseudocohnilembus species.     

Phylogenetic positions of two marine ciliates, Metanophrys similis and Pseudocohnilembus hargisi (Protozoa, Ciliophora, Scuticociliatia), inferred from complete small subunit rRNA gene sequences

The small subunit rRNA (SSrRNA) gene was sequenced for two marine scuticociliates Metanophrys similis and Pseudocohnilembus hargisi. The results show that this gene comprises 1763 and 1753 nucleotides in the two marine ciliates respectively. Metanophrys similis is phylogenetically closely related to the clade containing Mesanophrys carcini and Anophyroides haemophila, which branches basally to other species within the order Philasterida. Pseudocohnilembus hargisi groups with its congener, P. marinus, with strong bootstrap support. Paranophrys magna groups with the clade including Cohnilembus and Uronema, representing a sister clade to that containing the two Pseudocohnilembus species.     

Phylogenetic positions of two marine ciliates, Metanophrys similis and Pseudocohnilembus hargisi (Protozoa, Ciliophora, Scuticociliatia), inferred from complete small subunit rRNA gene sequences

The small subunit rRNA (SSrRNA) gene was sequenced for two marine scuticociliates Metanophrys similis and Pseudocohnilembus hargisi. The results show that this gene comprises 1763 and 1753 nucleotides in the two marine ciliates respectively. Metanophrys similis is phylogenetically closely related to the clade containing Mesanophrys carcini and Anophyroides haemophila, which branches basally to other species within the order Philasterida. Pseudocohnilembus hargisi groups with its congener, P. marinus, with strong bootstrap support. Paranophrys magna groups with the clade including Cohnilembus and Uronema, representing a sister clade to that containing the two Pseudocohnilembus species.     

丙酸酯变性淀粉浆料的研究

采用丙酸酐作为酯化剂,通过改变丙酸酐对玉米淀粉的投料比,制备一系列具有不同取代度(DS)的丙酸酯淀粉,以研究淀粉丙酸酯化变性程度与浆渡黏附性能和浆膜性能的关系,并通过与醋酸酯化变性的对比,来评价淀粉丙酸酯化淀粉的上浆性能.实验结果表明:淀粉丙酸酯化变性能够显著改善淀粉对涤纶纤维的黏附性能,提高淀粉浆膜的断裂伸长率和耐屈曲次数,降低浆膜的磨耗;随着取代度的增加,丙酸酯淀粉时涤纶纤维的黏附力先增大后减小,当取代度为0.019时,黏附力达到最大;相近取代度下,丙酸酯淀粉的浆膜性能及对涤纶纤维的黏附性能均优于醋酸酯淀粉.作为浆料使用的丙酸酯淀粉,其变性程度不宜过大.     

粗糙表面反射辐射偏振特性研究

为分析和研究粗糙表面反射辐射偏振特性及在目标表面二维空间的分布状况,基于偏振双向反射分布函数模型,推导出了粗糙表面反射辐射偏振度的一般表达式.针对不同材料涂层表面,综合考虑了表面漫反射和镜面反射,对偏振度进行了计算,分析了不同材料折射率及粗糙度下偏振度的分布.计算结果与实际测量数据基本吻合,为实际测量提供一定的理论指导.     

基于熵理论的高等学校教学管理系统评价模型

以熵理论为基础,分析了高校管理机构对系统内信息流的影响,然后从信息的角度对管理机构的有序度进行评价,引入了信息流时效和质量的概念,建立了管理机构优化设计和定量评价的时效质量模型,该模型可进行系统结构的有序度计算,并给出了效质有序度计算方法.     

互见中基于AIS数据的船舶领域

为确定船舶领域,根据琼州海峡船舶自动识别系统(AIS)数据,再现该水域内船舶航行过程.依据海上避碰规则,判断会遇船舶间避让义务关系,建立船舶避让行为和最近会遇点位置间的关系模型.利用数理统计和模糊数学方法,获得不同避让度下船舶领域的边界曲线,建立海上船舶领域模型.     

对形如(n~2+ln+k)~(1/2)的十分位问题的讨论

对于实数x,设d(x)是x的十进制表示中的十分位数。对正整数l和k的形如(n2+ln+k(l,n))~(1/2)=1取值进行研究,用初等方法,完整的讨论了取1,2,…,9时的可能性,及对应的n的范围。     

变性淀粉在赤铁矿阳离子反浮选脱硅中的抑制性能

以十二胺为捕收剂,木薯原淀粉、取代度为0.026和0.21的羧甲基淀粉和取代度为0.0065和0.055的磷酸酯淀粉作为抑制剂,考察了赤铁矿与石英的可浮性,重点研究了基团取代度对变性淀粉抑制性能的影响.结果表明:原淀粉、取代度0.026的羧甲基淀粉和取代度0.0065的磷酸酯淀粉对赤铁矿有良好的抑制作用,而取代度0.21的羧甲基淀粉和取代度0.055的磷酸酯淀粉对赤铁矿的抑制能力较弱;原淀粉和取代度0.026的羧甲基淀粉对石英有较强抑制作用,其他3种淀粉对石英抑制能力较弱.可见,低取代度的磷酸酯淀粉,在赤铁矿阳离子反浮选脱硅中可作为较高选择性的抑制剂.Zeta电位测定结果表明,特征基团取代度相对较高的变性淀粉,与赤铁矿和石英作用后,矿物Zeta电位负值较大.变性淀粉的取代度越高,其伸展向溶液中荷负电的基团越多,使阳离子捕收剂通过静电作用吸附于矿物表面,减弱了变性淀粉的抑制能力.     

关于模态综合法模态函数选择和计算精度的研究

为了提高模态综合法的综合效率和计算精度,对系统分部的划分、模态函数的选择和系统自由度数的确定作了研究.并重点放在选用幂级数作为分部形态函数时对系统计算精度的影响方面.     

两种不同混凝土异形柱正截面极限承载力的比较

提出钢骨混凝土异形柱的概念,通过4根不对称T形截面钢骨混凝土异形柱正截面承载力的试验研究,揭示了钢骨混凝土异形柱的工作机理、破坏形态和极限承载力,验证了平均应变的平截面假定.编写钢骨混凝土异形柱正截面承载力计算的计算机程序.试验和程序计算结果表明,钢骨混凝土异形柱与钢筋混凝土异形柱相比,承载力明显提高.     

微波法制备淀粉醋酸酯的研究

以淀粉为原料,冰醋酸和醋酸酐为改性剂,在微波加热条件下制备淀粉醋酸酯。探讨了微波火力、微波加热时间、淀粉种类及醋酸酐用量对淀粉醋酸酯取代度和反应效率的影响。结果表明:当玉米淀粉:冰醋酸:醋酸酐的质量分数比为1∶1∶0.4时,在中火条件下微波加热6 min,制得的淀粉醋酸酯的取代度为0.5386,反应效率为84.78%;在实验范围内,随着醋酸酐加入量的增加,制得的淀粉醋酸酯的取代度不断增加,而反应效率呈下降趋势;在相同的条件下,木薯淀粉制得的淀粉醋酸酯取代度和反应效率高,玉米淀粉次之,马铃薯淀粉制得的淀粉醋酸酯取代度和反应效率较低。     

The composition and distribution of metal clusters in the MoFe protein from a nifZ deletion strain （DJ 194） of Azotobacter vinelandii

Through the anaerobic chromatography on the columns of DEAE 52, Q-Sepharose and Sephacryl S-200, a nitrogenase MoFe protein (△nifZ Av1) was obtained from a nifZ deleted mutant of Azotobacter vinelandii (stain DJ194).The results of Western blotting after anoxic native electrophoresis and SDS-PAGE showed that △nifZ Av1 was similar to wild type MoFe protein (OP Av1) at the electrophoretic mobility, molecular weight and subunit composition. Furthermore, △nifZ Avl was also similar to OP Av1 at the molybdenum content, EPR signal (g≈4.3, 3.65 and 2.01), and the molar extinction coefficient (△ε) of circular dichroism (CD)at 660 nm region. All of these indicated that, besides having the same α2β2 composition as OP Av1, the △nifZ Av1 also contained equal amount of reductive FeMoco in the spin state of S=3/2 to OP Av1. However, the iron content and substrate (C2H2, H^ and N2)-reduction activity of △nifZ Av1 were 74% and 46%-50% of those of OP Av1, respectively. Furthermore, the △ε at around 450 nm, which reflects P-cluster in Av1, was obviously lower than that of OP Av1. It suggested that the difference between △nifZ Avl and OP Av1 resulted from P-cluster rather than FeMoco, and from the half number of P-cluster in △nifZ Av1, but the composition or redoxstate of P-cluster in △nifZ Av1 were not changed. Thus it could propose that △nifZ Av1 is composed of two different αβsubunit pairs. One is a FeMoco-and P-cluster-containing pair, and the other is a P-cluster-deficient but FeMoco-containing pair. Since the deletion of nifZ gene leads to the deficiency of only one of two P-clusters in a α2β2 tetramer, the assembly of P-cluster may not simply depend on one gene product, and so a possible mechanism of NifZ is supposed here.