定向进化提高灰盖鬼伞过氧化物酶染织废水脱色效率

【目的】获得染织废水脱色能力增强的灰盖鬼伞过氧化物酶。【方法】使用基因合成及定点突变平台合成突变灰盖鬼伞过氧化物酶基因CIPmt4(I49S、V53A、M166F和M242I),并调整密码子至毕赤酵母偏好性。以CIPmt4为模板进行定向进化,经过三轮易错PCR和高通量筛选得到一个酶学性质显著改善的突变体(CIPmt5)。通过3D建模和分子动力学模拟分析蛋白的结构及热稳定性,并进一步研究CIPmt5和野生型CIP对刚果红、氨基黑、甲基橙、次甲基蓝、苯胺蓝、结晶紫、溴酚蓝共7种染料的脱色能力。【结果】序列分析显示该突变体积累了I49S、V53A、T121A、M166F和Y272F共5个氨基酸突变,与野生型灰盖鬼伞过氧化物酶相比,以ABTS为底物酶活性是野生型的2.01倍(24.44 U/mg),最适反应p H由5.0提高到6.5,最适反应温度由25°C提高到45°C。除次甲基蓝外对其它染料脱色的最适p H都往中、碱性方向偏移,脱色率普遍高于野生型。模型分析显示CIPmt5活性中心更开放,热稳定性增强。【结论】突变体酶CIPmt5能够更好地替代野生型灰盖鬼伞过氧化物酶应用于染织业染料脱色、化工废水和染织废水的生物修复。     

重金属污染环境的植物修复及其分子机制

重金属污染物的排放和扩散造成了日益严重的环境污染。如何消除环境中的重金属污染物已成为国际性难题。近年来,植物修复技术的出现和快速发展为我们展示了一条新的有效途径：即利用植物对重金属化合物的吸收、富集和转化能力去除土壤和水体中残存的重金属污染物。文章简要介绍了重金属污染物与植物修复的关系和植物修复的生理机制,重点总结了重金属污染环境的植物修复在分子生物学方面所取得的研究进展,包括有机汞裂解酶基因merB、汞离子还原酶基因merA和金属硫蛋白基因MT等的生物学功能及其在植物修复上的应用,展望了植物修复研究工作的发展方向,并针对汞污染提出了一套修复方案。     

水稻双链RNA结合蛋白同源基因OsRBP的克隆及其表达的分析

在基因数据库中发现两个水稻EST片段与大白菜BcpLH基因的双链RNA结合结构域 (dsRBD)有同源的区域 ,根据同源片段的位置特征设计引物 ,用RT-RCR扩增粳稻 (Oryzasativasubsp .japonica)愈伤组织的cDNA ,得到大小为 1.8kb的DNA片段。该cDNA片段含完整的编码区 ,有两个典型的dsRBD ,分别与BcpLH的dsRBD在氨基酸水平上同源性为 75 %左右 ,故将其命名为OsRBP。RT -PCR表达分析显示该基因在未成熟的种子和愈伤组织中表达 ,在根、茎、叶、穗、成熟种子及胚芽鞘中没有表达信号 ,由此推测该基因的表达可能与种子和胚的早期发育相关。该研究首次从水稻中分离到双链RNA结合蛋白基因 ,并初步研究了其表达方式 ,为进一步探讨水稻重要器官的发育和植物中双链RNA结合蛋白的调节作用奠定了基础     

Human alpha-galactosidase A: characterization of the N-linked oligosaccharides on the intracellular and secreted glycoforms overexpressed by Chinese hamster ovary cells

Human alpha-galactosidase A (alpha-Gal A) is the lysosomal glycohydrolase that cleaves the terminal alpha-galactosyl moieties of various glycoconjugates. Overexpression of the enzyme in Chinese hamster ovary (CHO) cells results in high intracellular enzyme accumulation and the selective secretion of active enzyme. Structural analysis of the N -linked oligosaccharides of the intracellular and secreted glycoforms revealed that the secreted enzyme's oligosaccharides were remarkably heterogeneous, having high mannose (63%), complex (30%), and hybrid (5%) structures. The major high mannose oligosaccharides were Man5-7GlcNAc2 species. Approximately 40% of the high mannose and 30% of the hybrid oligosaccharides had phosphate monoester groups. The complex oligosaccharides were mono-, bi- , 2,4-tri-, 2,6-tri- and tetraantennary with or without core-region fucose, many of which had incomplete outer chains. Approximately 30% of the complex oligosaccharides were mono- or disialylated. Sialic acids were mostly N -acetylneuraminic acid and occurred exclusively in alpha2, 3-linkage. In contrast, the intracellular enzyme had only small amounts of complex chains (7.7%) and had predominantly high mannose oligosaccharides (92%), mostly Man5GlcNAc2 and smaller species, of which only 3% were phosphorylated. The complex oligosaccharides were fucosylated and had the same antennary structures as the secreted enzyme. Although most had mature outer chains, none were sialylated. Thus, the overexpression of human alpha-Gal A in CHO cells resulted in different oligosaccharide structures on the secreted and intracellular glycoforms, the highly heterogeneous secreted forms presumably due to the high level expression and impaired glycosylation in the trans- Golgi network, and the predominately Man5-7GlcNAc2 cellular glycoforms resulting from carbohydrate trimming in the lysosome.      

Histological and immunohistochemical studies of the structure of lymph nodes in Kilis goats

Ten healthy adult Kilis goat mesenteric lymph nodes were used to examine the general structure of lymph nodes, lymphocytes, plasma cells, reticular cells and reticular fibers using histological methods. We also detected T lymphocytes using anti-CD3 [SP7], anti-CD4 [74-12-4], mouse anti-bovine CD4 [CC30] and mouse anti-bovine CD8 [CC63] monoclonal antibodies (mAb); and B lymphocytes using anti-CD79a [HM57] mAb, macrophages using anti-macrophage [MAC387] mAb and follicular dendritic cells using anti-S100 polyclonal antibody (pAb). The distribution of these cells also was studied. Although the primer antibodies we used for CD3, CD8, CD79a, MAC387 and S100 worked well, the primer antibodies for CD4 were ineffective for paraffin embedded goat lymph nodes.     

Quantitative trait loci analysis of individual and total isoflavone contents in soybean seeds

Soybean isoflavones play diverse roles in human health, including cancers, osteoporosis, heart disease, menopausal symptoms and pabulums. The objective of this study was to identify the quantitative trait loci (QTL) associated with the isoflavones daidzein (DC), genistein (GeC), glycitein (GlC) and total isoflavone contents (TIC) in soybean seeds. A population of 184 F_21:0 recombinant inbred lines derived from a ‘Xiaoheidou’ בGR8836’ cross was planted in pot and field conditions to evaluate soybean isoflavones. Twenty-one QTL were detected by composite interval mapping. Several QTL were associated with the traits for DC, GeC, GlC and TIC only. QDGeGlTIC4_1 and QDGlTIC12_1 are reported first in this study and were associated with the DC, GeC, GlC and TIC traits simultaneously. The QTL identified have potential value for marker-assisted selection to develop soybean varieties with desirable isoflavone content.     

榕树须活性部位的成分研究

采用硅胶柱层析色谱和重结晶方法对榕树须95%乙醇提取物进行分离,得到8化合物,通过理化性质和波谱技术分别鉴定为:α-香树脂醇乙酸酯(1),羽扇豆醇乙酸酯(2),木栓酮(3),木栓醇(4),正三十五烷醇(5),白桦酯酸(6),β-谷甾醇(7)和豆甾-4-烯-6β-羟基-3-酮(8)。除化合物6外,其余化合物均为首次从该植物中分离得到。     

一个重要的蒙古二叠纪牙形刺种

在蒙古南部Lugiingol剖面卢金格尔(Lugiingol)组下部发现牙形刺Mesogondolell aidahoensis,该地层应归入下二叠统空谷阶,而不是像以前那样归入上二叠统。