Development of an efficient process intensification strategy for enhancing <Emphasis Type="Italic">Pfu</Emphasis> DNA polymerase production in recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis>

An efficient induction strategy that consisted of multiple additions of small doses of isopropyl-β-D-thiogalactopyranoside (IPTG) in the early cell growth phase was developed for enhancing Pfu DNA polymerase production in Escherichia coli. In comparison to the most commonly used method of a single induction of 1 mM IPTG, the promising induction strategy resulted in an increase in the Pfu activity of 13.5 % in shake flasks, while simultaneously decreasing the dose of IPTG by nearly half. An analysis of the intracellular IPTG concentrations indicated that the cells need to maintain an optimum intracellular IPTG concentration after 6 h for efficient Pfu DNA polymerase production. A significant increase in the Pfu DNA polymerase activity of 31.5 % under the controlled dissolved oxygen concentration of 30 % in a 5 L fermentor was achieved using the multiple IPTG induction strategy in comparison with the single IPTG induction. The induction strategy using multiple inputs of IPTG also avoided over accumulation of IPTG and reduced the cost of Pfu DNA polymerase production.     

Two-Photon Correlation Spectroscopy in Single Dendritic Spines Reveals Fast Actin Filament Reorganization during Activity-Dependent Growth

Two-photon fluorescence correlation spectroscopy (2P-FCS) within single dendritic spines of living hippocampal pyramidal neurons was used to resolve various subpopulations of mobile F-actin during activity-dependent structural changes such as potentiation induced spine head growth. Two major classes of mobile F-actin were discovered: very dynamic and about a hundred times less dynamic F-actin. Spine head enlargement upon application of Tetraethylammonium (TEA), a protocol previously used for the chemical induction of long-term potentiation (cLTP) strictly correlated to changes in the dynamics and filament numbers in the different actin filament fractions. Our observations suggest that spine enlargement is governed by a mechanism in which longer filaments are first cut into smaller filaments that cooperate with the second, increasingly dynamic shorter actin filament population to quickly reorganize and expand the actin cytoskeleton within the spine head. This process would allow a fast and efficient spine head enlargement using a major fraction of the actin filament population that was already present before spine head growth.     

夏枯草DNA提取及RAPD反应条件的优化

用改进后的SDS法从夏枯草植物的新鲜叶片中提取基因组DNA,通过正交设计和单因素结合的方法对RAPD-PCR反应条件进行优化,确定了适合夏枯草DNA的最佳扩增体系:20μL的PCR反应体系中,模板DNA浓度1.0-2.0 mg/L,引物浓度1.00-1.50 mmol/L,Mg2 浓度2.0-2.5 mmol/L,dNTP浓度0.20-0.25 mmol/L,Taq酶用量0.5 U,10×BufferMg2 free 2μL,BSA浓度0.25-0.50μg/μL。     

棘胸蛙、虎纹蛙、黑斑蛙血细胞的比较

应用Giemsa染色法对棘胸蛙、虎纹蛙和黑斑蛙的血细胞进行显微观察和各形态参数测定,并对各类细胞作了图示和描述。结果表明,红细胞和白细胞的形态和数量比例有一定属和种的特征,并与其生长环境和进化地位有一定联系。     

四川动物体内发现的钩口线虫

报道寄生于四川17种动物体内的钩口线虫12种,它们隶属于钩口科的6个属,亦记述了部份宿主的感染情况。     

A Study on Karyotypes of 6 Species of Elymus in China

The present paper deals with the karyotype analysis of 6 species of Elymus which are native to Qinghai-Tibetan plateau. The number of somatic chromosomes in roottip cells of the 6 species and their karyotypes are reportad here for the first time, and they are all hexaploid, with 2n=6x=42. The karyotype formulae are as follows: E. melantherus, 2n=6x=42=32m+10sm, E. kengii, 2n=6x=42=34m+8sm, E grandiglumis, 2n=6x=42= 30 m+12 sm, E. laxiflorus, 2n=6x=42=32 m +10 sm, E. kokonoricus, 2n+6x=42=34 m +8 sm, E. longiglumis, 2n+6x=42=34 m+8 sm. No satellites have been founded in the 6 species and all the karyotypes belong to 1B or 2B type:     

Effect of Changes of Temperature Around Roots in Relation to Water Uptake by Roots and Leaf Transpiration

Effects of changes in temperature around roots on water uptake by roots and leaf transpiration were studied in Leucaena leucocephala (Lam.) de Wit., a subtropical woody plant species, and in Zea mays L. When the temperature around roots was rapidly lowered from 25 ℃ to 15 ℃, the water uptake by the roots and leaf transpiration were stimulated significantly within a short period ( 14 min). However, this effect did not occur when the cooling time was prolonged neither did if occur when the temperature around the roots was resumed from 15 ℃ to 25 ℃. Both the hydraulic conductivity of roots and leaf transpiration were increased substantially at first (within 20 min)and then decreased steadily to a level lower than those of the control in which the roots were continuous exposed to a low temperature ( 15 ℃ ). Low temperature also promoted the biosynthesis of ABA in roots and enhanced the xylem ABA concentration, but such stimulation did not occur untill about 30 min after cooling treatment, leaf transpiration was reduced markedly, but the hydraulic conductivity of roots increased when the root system was treated with exogenous ABA. It was suggested that some mechanisms other than ABA may be involved in the short-time cryostimulation of water uptake by roots and leaf transpiration.     

脊髓蛋白激酶Cα和γ亚型在吗啡依赖和戒断反应中的不同作用

在大鼠吗啡依赖和戒断模型上,采用行为学、免疫组织化学和Western blot方法观察鞘内应用蛋白激酶C(protien kinase C,PKC)抑制剂chelerythrine chloride(CHE)对吗啡依赖大鼠纳洛酮催促成断反应、脊髓Fos蛋白表达和脊髓神经元胞膜和胞浆PKCα、γ表达的影响,以探讨不同亚型PKC在吗啡依赖和戒断反应中的作用。结果表明,鞘内注射CHE能明显减轻吗啡成断症状的评分和吗啡戒断引起的痛觉异常,抑制吗啡成断期间脊髓Fos蛋白表达的增加;吗啡依赖可引起脊髓神经元PKCα和γ表达的上调和转位：吗啡戒断期间存在明显的且可被鞘内注射CHE抑制的PKCα转位,但未观察到明显的PKCγ转位。上述结果表明,脊髓PKC表达上调和转何可能参与吗啡依赖的形成和戒断反应的表达,且PKCα和γ亚型在吗啡依赖和戒断反应中的作用存在差异。     

鄂尔多斯高原油蒿种群分布格局对降水梯度的反应

应用方差均值比率法、聚块性指数及点格局方法,沿着鄂尔多斯高原从东至西的降水梯度(336～249 mm·a^-1),设置5个采样点,对油蒿种群分布格局及其对降水梯度的反应进行了研究.结果表明：随着降水梯度的递减,在小尺度上研究区油蒿种群分布格局表现为由均匀分布向随机分布转变;在大尺度上则表现为由随机分布向聚集分布转变.降雨的减少显著改变了油蒿种群的空间分布格局,在进行生态恢复时需对植物个体进行合理配置.     

郑汴路路旁土壤-小麦系统重金属积累及其健康风险评价

按离路基不同距离采集土壤、麦苗和籽粒样品, 在测定样品重金属 (Pb、Cd、Zn、Cr和Cu) 的基础上, 开展了路旁土壤-小麦系统重金属分布、积累和污染状况分析, 并对膳食小麦 (Triticumaestivum) 引起的健康风险进行了评价。结果表明:1) 土壤-小麦系统重金属含量随着离开路基距离的增加呈先增加后减少的趋势, 土壤重金属含量>麦苗重金属含量>籽粒重金属含量。2) 麦苗和小麦籽粒对土壤重金属富集能力的大小顺序均为Cu>Cd>Zn>Pb>Cr, 麦苗对重金属的富集能力大于小麦籽粒。3) 膳食小麦所致的Cd个人健康风险较大。