犬细小病毒VP2蛋白在真核细胞中的分泌表达及特性

摘要:【目的】利用真核细胞分泌表达犬细小病毒VP2蛋白和研究其特性。【方法】为构建犬细小病毒（Canine parvovirus, CPV）VP2基因的真核分泌型表达载体,首先通过酶切从含有人CD5信号肽序列的质粒中将CD5信号肽基因片段切出,将其连接到真核表达载体pcDNA3.1A的多克隆位点上,构建成pcDNA3.1-CD5sp质粒。然后再通过PCR方法从含有犬细小病毒VP2基因的质粒中扩增VP2基因,并将其插入到pcDNA3.1- CD5sp载体中CD5信号肽的下游,构建成VP2基因的真核分泌型表达载体pcDNA-CD5sp-VP2。经磷酸钙介导转染293T细胞,使其在真核细胞中进行分泌表达,并通过ELISA检测表达的VP2蛋白与犬转铁蛋白受体（TfR）结合的活性。【结果】序列分析结果表明,本实验构建的犬细小病毒VP2基因真核分泌型表达载体结构正确,将该表达载体转染的293T细胞,在培养基中通过Western-blot检测到有VP2重组蛋白的存在。经ELISA检测表明表达的重组VP2蛋白具有与犬转铁蛋白受体结合的活性。【结论】 利用人的CD5信号肽实现了犬细小病毒VP2蛋白在真核细胞中的分泌表达,表达的VP2蛋白具有与犬转铁蛋白受体结合的活性。     

一株种间融合的三倍体杂种酵母减数分裂产物的遗传学分析

通过原生质体融合技术将二倍体酿酒酵母（Ｓａｃｃｈａｒｏｍｙｃｅｓｃｅｒｅｖｉｓｉａｅｖａｒ．ｅｌｌｏｐｓｉｏｄｅｕｓ）与单倍体糖化酵母（Ｓａｃｃｈａｒｏｍｙｃｅｓｄｉａｓｔａｔｉｃｕｓ）构建成遗传上稳定的种间三倍体融合杂种。实验结果表明,这种融合杂种象有性杂种一样能诱导产孢;对其完整和非完整四分子的遗传分析,证明了通过遗传标记互补选择法获得的种间三倍体融合杂种ＨＵ－ＫＤＦ－２４０在产孢过程中,标记基因发生了分离和交换,出现了亲二型和重组类型;四分子对可溶性淀粉的发酵和不发酵分离比为１∶２,而原养型与营养缺陷型的分离比是１∶１。     

冬小麦种植模式对水分利用效率的影响

在同一种植密度下,设3种种植模式,包括25 cm等行距平作、"20+40"大小行平作和"20+40"沟播。研究了冬小麦沟播和平作种植对产量及水分利用效率的影响。结果表明,"20+40"沟播产量显著高于平作;叶片相对含水量(RWC)、水势(Ψw)和叶片水平水分利用效率随生育进程的推进呈整体下降趋势,其中,沟播处理RWC、Ψw和叶片水平水分利用效率的平均值均显著高于等行距和"20+40"平作处理;另外,"20+40"沟播还能明显提高冬小麦田土壤贮水量,减少总耗水量,从而提高水分利用效率。灌水增加了冬小麦产量和叶片相对含水量等各水分指标,降低了水分利用效率,减小了各种植模式间差异。"20+40"沟播在灌水135 mm条件下既保障产量又较等行距节水25%。由此表明,冬小麦"20+40"沟播可改善叶片水分状况,提高水分利用效率,增加作物产量。     

内蒙古半干旱生态脆弱矿区生态修复耦合机理与产业模式

近年,煤炭开采带动矿区经济发展的同时也造成了严重的生态环境问题。由此产生生态修复和土地复垦产业延伸的生态环境治理和矿区产业调整优化方面研究热点,二者形成的耦合系统可在恢复受损土地的同时,以治理和循环生产为契机延伸产业链。受损地形重塑及边开采边恢复技术开发及应用形成安全稳定地形条件是前提,其次是土壤的恢复重构,在当地条件下迅速恢复具备良好理化性质,供植被和经济能源作物生长和景观格局构建,最终形成修复产业和生态产业的价值创造平台。根据耦合系统的工艺特点,每年内蒙矿区利用该耦合系统以矿区受损土地修复进行循环生产生态修复产品的潜力约21915.51万吨和经济能源作物的潜力约21.15万t。     

鱼类基因内含子研究进展

内含子是指断裂基因中的非编码区序列,在编码蛋白质前被去除。在高等生物中,内含子的长度远大于外显子,大部分随机突变会发生在内含子中。因此,内含子的存在使高等生物对突变的耐受能力大大增强了。研究表明,内含子可以提高基因表达效率;影响RNA的转录、剪接加工、出核孔以及翻译等过程;启动某些基因的表达;并通过选择性剪接调控基因的表达。内含子功能的研究成果给当前鱼类免疫基因研究开拓了全新的视野。对内含子的分类、剪接、功能以及鱼类内含子研究的新进展进行了综述,并展望了内含子在鱼类免疫基因研究中的应用。     

中国南海北部不同岛屿植被与土壤的耦合协调发展度

植被和土壤是陆地生态系统两个重要的组成部分,二者相互影响相互促进,探明两者的耦合协调关系是生态恢复与重建的顺利实施的关键。基于中国南海北部大庙墩岛、涠洲岛、大汉三墩岛、甘蔗岛和蜈支洲岛等个海岛的典型植被群落植被土壤的全面调查和取样分析,建立10个植被因子和14个土壤因子的2级层次指标体系,采用层析分析法确定各因子的权重,构建5个海岛植被土壤耦合度和耦合协调度模型。结果表明,不同岛屿的植被土壤耦合度和耦合协调度模型并不完全对应,植被与土壤的综合指数在不同岛屿中也不完全一致,甘蔗岛的植被效应和大汉三墩岛的土壤效应最佳。中国南海北部5个岛屿的植被土壤耦合协调状况较好,均处于初、中级协调发展状态,且除甘蔗岛外处于植被土壤同步型。总的来说,由于岛屿远离内陆,人类干扰相对较小,在植被土壤长期的演替过程中,中国南海北部岛屿植被土壤耦合协调较好,在其恢复与生态重建时要注重提高植物多样性、抚育水平和土壤管理水平。     

Scutellarin ameliorates nonalcoholic fatty liver disease through the PPARγ/PGC-1α-Nrf2 pathway

Nonalcoholic fatty liver disease (NAFLD) is characterised by excessive accumulation of hepatic lipids and oxidative injury of hepatocytes. Scutellarin is a flavonoid glycoside having antioxidative stress activity. Our current study aims to investigate the molecular mechanism of scutellarin ameliorating NAFLD. Scutellarin treatment was applied to male C57BL/6 mice maintained on a high-fat diet (HFD) and HepG2 cells challenged with oleic acid. The antioxidation biochemical indicators and lipid levels in the liver and cells were detected by kits. Liver pathology was observed by light microscope, Oil Red O staining, and transmission electron microscope (TEM). In addition, quantitative real-time polymerase chain reactions (qRT-PCR) and western blot assays were employed to detect the mRNA and protein levels of various antioxidative-related genes in the presence or absence of peroxisome proliferator-activated receptor gamma (PPARγ); inhibitor GW9662. Our results showed that scutellarin could significantly reduce blood lipid levels and enhance antioxidative capacities in both the models. In addition, scutellarin treatment conspicuously activated PPARγ, peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α), nuclear factor erythroid-2-related factor (Nrf2), haem oxygenase-1 (HO-1), glutathione S-transferase (GST), and NAD(P)H quinone dehydrogenase one (NQO1), while it significantly inhibited nuclear factor kappa B (NF-κB), Kelch-like ECH-associated protein 1 (Keap1) at both the mRNA and protein levels. However, after interfered by GW9662, scutellarin effect was significantly decreased. The experimental data demonstrated that scutellarin showed strong hypolipidaemic, antioxidative, and liver protective activity which could be attributed to its regulating activity in the PPARγ/PGC-1α-Nrf2 signaling pathway.     

Heavy metal contamination and health risk assessment in soil-rice system near Xinqiao mine in Tongling city,Anhui province,China

In this study, paddy soil and rice grain samples were collected from the vicinity around the Xinqiao mine in Tongling, China to test for the presence of heavy metals (Cd, Ni, Cr, Cu, Zn, and Pb) in soil-rice system. Results indicated that the soil samples were primarily contaminated with Cd and Cu and followed with Zn and Pb. In rice grains, Cd, Cu, and Cr concentrations exceeded recommended guidelines. However, the regional distribution of heavy metals in rice grains and soil was inconsistent. The bioaccumulation factor of heavy metals in rice grains decreased in the order of Cd > Zn > Cu > Ni > Cr > Pb. The BAF was significantly positively correlated with TCLP-extractable metals and significantly negatively correlated with soil pH. However, the relationship between soil organic matter and the BAF in rice grains was complex. Health risk assessment through rice intake showed that hazard quotients of Cu and Cd were greater than 1 and could pose a considerable non-cancer health risk to adults and children; meanwhile, Cr, Ni, and Cd could pose an unacceptable cancer risk. The results indicated that the government must take measures to reduce heavy metal contents in paddy soil and rice.     

Novel inhibitors of As(III) S-adenosylmethionine methyltransferase (AS3MT) identified by virtual screening

Due to increased interest in As(III) S-adenosylmethionine methyltransferase (AS3MT), a search for chemical probes that can help elucidate function was initiated. A homology model was built based on related enzymes, and virtual screening produced 426 potential hits. Evaluation of these compounds in a functional enzymatic assay revealed several modest inhibitors including an O-substituted 2-amino-3-cyano indole scaffold. Two iterations of near neighbor searches revealed compound 5 as a potent inhibitor of AS3MT with good selectivity over representative methyltransferases DOT1L and NSD2 as well as a representative set of diverse receptors. Compound 5 should prove to be a useful tool to investigate the role of AS3MT and a potential starting point for further optimization.