超嗜热酯酶EST2在不同宿主中的异源高效表达研究

来源于超嗜热古菌Alicyclobacillus acidocaldarius的酯酶EST2是目前报道的活性最高的超嗜热酯酶,具有极大的工业应用价值。为促进EST2的生产应用,将其分别在大肠杆菌及毕赤酵母中进行异源表达,并就不同宿主对表达情况和重组酶酶学性质的影响进行了分析。在大肠杆菌和毕赤酵母中重组表达的EST2酶学性质基本一致:最适温度分别为75℃和77.5℃,最适pH均为8.0,比活力分别为4656.6 U/mg和4078.3 U/mg,70℃水浴保温4.5 h,残余活力均在70%以上。在摇瓶发酵的基础上,于5 L发酵罐中进行了重组大肠杆菌及毕赤酵母的高密度发酵。毕赤酵母高密度发酵120 h菌体干重达68 g/L,最大表达酶活力为959.6 U/ml。大肠杆菌高密度发酵25 h菌体干重达60.8 g/L,最大酶活力14825.6 U/ml,表达量是毕赤酵母的15.4倍,单位时间产量是酵母的74.2倍。结果表明大肠杆菌发酵周期短、表达量高,更适合进行嗜热酯酶EST2的高效生产,这为促进嗜热酯酶在工业生物技术产业的应用奠定了基础。     

中国灵芝科真菌资源与分布

中国灵芝科资源十分丰富。许多种类具有重要的经济价值。现在已知灵芝种类有88种。本文概述了灵芝对国民经济的重要性,中国古籍中有关灵芝的记载,灵芝的化学成分与制剂,灵芝的临床应用,灵芝的人工培养,灵芝的生态习性与分布。     

Production and Accumulation of Xylooligosaccharides with Long Chains by Growing Culture and Xylanase of a Mutant Strain of Bacillus pumilus X-6-19

Bacillus pumilus X-6-9 isolated from soil and subsequently identified, produced xylooligosacchatides with long chainsfrom xylan and accumulated them in the culture. By improving the culture conditions and mutating the bacterium, a 3.2-fold increasein the production of the xylooligosaccharides was established, when compared to the original culture conditions of B. pumilus X-6-19.The addition of D-glucose to the culture of the mutant swain U-3 of B. pumilus X-6-9 repressed the synthesis of β-xylosidase, but notxylanase. Thus, it was revealed that strain U-3 was a good organism for the production and accumulation of xylooligosaccharideswith long chains from xylan by a microbial culture. Xylanase produced by strain U-3 was purified to homogeneity and characterized.The hydrolyzates generated by the purified xylanase contained xylobiose, xylotrinse, xylotewaose, and xylopentaose, but not xylose.     

土壤有机磷研究进展

土壤有机磷在农业生态系统的物质循环过程中具有重要作用．研究土壤有机磷,对深入了解土壤-植物系统中磷的营养循环特征具有重要意义．结合国内外已有成果和最新研究进展,从土壤有机磷含量及其影响因素、土壤有机磷的形态和测定方法、土壤有机磷的分组和施肥的影响、土壤有机磷的矿化等几个方面综述了国内外土壤有机磷的研究现状,并提出了目前在土壤有机磷研究中存在的一些问题以及今后研究的热点．     

锦鲤4个人工雌核发育家系的微卫星标记研究

利用Crooijmans et al.(1997)分离的包含CA重复单元的普通鲤鱼（Cyprinus carpio L.)的8个微卫星DNA标记,对从锦鲤（Cyprinus carpio L.)的红白,大正和昭和3个不同品系中所获得的4个不同人工雌核发育家系的20尾个体进行PCR扩增。电泳结果表明,8对引物在20尾个体中均能重复稳定地扩增出相应的同源序列。随引物不同,各等位基因数为1－11个,大小在68－264bp。在MFW4,MFW7,MFW19,MFW20,MFW23和MFW24 6个微卫星的扩增结果中,20尾个体的扩增图谱呈现了高度的遗传多态性,不同雌核发育家系内个体的遗传异质性也较大。其中大正（TaS)和红白1（RW1）的个体不仅花色分化显著,而且个体间的平均遗传距离分别高达0．28。通过对微卫星等位基因和基因型分析发现,由于锦鲤品系中的每一个体是通过不断地杂交选育而获得,基因组来源复杂,基因高度杂合。因此,只进行1代的人工雌核发育,其家系内仅部分个体的部分座位出现纯合。所获得的人工雌核发育锦鲤为后续的色素遗传调控机制研究提供了必要的实验材料;同时,所鉴定的微卫星分子标记为进行锦鲤的分子标记育种的基因组作图提供了理想的工具。     

PNAS-4 expression and its relationship to p53 in colorectal cancer

PNAS-4 is a novel pro-apoptotic protein activated during the early response to DNA damage; however, the molecular mechanisms and pathways regulating PNAS-4 expression in tumors are not well understood. We hypothesized that PNAS-4 is a p53 down-stream target gene and designed this study. We searched online for putative p53-binding sites in the entire PNAS-4 gene and did not find any corresponding information. In HCT116 colon cancer cells, after being transfected with small interfering RNA to silence p53, the expressions of PNAS-4 and other known p53 target gene (Apaf1, Bax, Fas and Dr5) were determined by real-time PCR. We found that PNAS-4 was up-regulated while Apaf1, Bax, Fas and Dr5 were down-regulated. We then examined the expression of PNAS-4 and p53 mutation in colorectal cancer patients. PNAS-4 expressed both in colorectal cancers and normal tissues, but compared with paired control, PNAS-4 was up-regulated in cancers (P = 0.018). PNAS-4 overexpression ratios were correlated to the p53 mutant status (P = 0.001). The mean PNAS-4 expression levels of p53 mutant homozygote group and heterozygote group were higher than that of p53 wild type group (P = 0.013). The expression ratios of PNAS-4 (every sample in relative to its paired normal mucosa) were different between negative lymph node metastasis (66% up-regulated, 34% down-regulated) and positive metastasis (42% up-regulated, 58% down-regulated). Taken together, these findings suggested that PNAS-4 was not a p53 target, but overexpression of PNAS-4 was correlated to p53 inactivity in colorectal cancer.     

长柄水青冈幼苗根系构型及动态变化

长柄水青冈（Fagus longipetiolata）是我国亚热带山地林的重要组成树种,本研究在温室中用河砂培养长柄水青冈幼苗,采用直接测量法研究了长柄水青冈的根系构型。结果表明,生长50 d的长柄水青冈幼苗的根系呈倒圆锥形,根宽小于根深;基根平均生长角度较小,基根的向地性小;一级侧根与主根的夹角从根系上部至下部逐渐变小;二级侧根首先发生于根系的中下部,然后其发生范围向根系上、下部扩展。虽然长柄水青冈幼苗根系构型存在较大的个体差异性,但一级侧根与主根的夹角则具有较好的稳定性。研究揭示了实验条件下长柄水青冈幼苗的根系构型及其在幼苗建成过程中的生长变化规律,为长柄水青冈的进一步研究提供科学依据。     

胶质芽胞杆菌对黑麦草生长特性的影响

研究了从玉米(K02)、草地早熟禾(K05)、披碱草(K09)、多年生黑麦草(K11)、匍匐翦股颖(K12)根际土壤中分离到的5株胶质芽胞杆菌菌株对黑麦草株高、叶绿素含量以及根系活力的影响。结果表明在苗期(播种后10 d)、中期(播种后50 d)以及收获期(播种后90 d)3个采样期各处理黑麦草株高、叶绿素含量和根系活力均高于对照(CK)。研究表明施入胶质芽胞杆菌菌剂对黑麦草株高、叶绿素含量和根系活力具有一定的促进作用,其结果可为生物钾肥的研制提供必要的数据。     

基于遥感和GIS的川西绿被时空变化研究

探索出了利用多时相MODIS数据分析绿被时空变化的技术方法,揭示出了川西2002—2008年间绿被的变化特征。建立了绿被提取模型。用其从2002—2008年的多期MODIS影像中提取出川西多期绿被数据。其次,利用GIS技术对2002—2008年间绿被变化,及其与温度、降雨的关系等进行了分析。研究表明:2002年绿被天数在195 d以上的区域有43.3%。终年无绿被的区域有4.3%。西北部植被生长日数短,而东南部生长日数长。2002—2008年间,春季绿被面积变幅最大,秋季最小。最大绿被面积出现在2006年的夏季,最小绿被面积出现在2005年的冬季。平均夏冬绿被面积差占区域面积46.7%。在季节上,绿被面积与温度和降雨量均在0.01的水平上呈显著正相关关系,其相关系数分别为0.82和0.84。该研究成果对植被生长潜力挖掘、农牧生产和生态建设决策等均有重要意义。