大鼠Bdnf基因慢病毒载体的构建及其在骨髓间质干细胞中的表达

骨髓间质干细胞(MSCs)是目前基因工程正在探讨应用的靶细胞,为构建带有脑源性神经营养因子(Bdnf)基因慢病毒载体并使其在大鼠骨髓间质干细胞中表达,采用RT-PCR技术获得大鼠Bdnf基因编码区(CDS)片段,限制性内切酶酶切和基因重组构建慢病毒载体质粒PNL-BDNF-IRES2-EGFP,在脂质体介导下与包装质粒HELPER,包膜质粒VSVG共转染293T细胞包装生产慢病毒。所获慢病毒感染大鼠MSCs(rMSCs)后,PCR和免疫细胞化学法检测在rMSCs中Bdnf基因的插入和表达。结果显示所获的Bdnf基因经测序后与GenBank报道序列完全一致。重组慢病毒载体质粒PNL-BDNF-IRES2-EGFP经鉴定正确。三质粒共转染293T细胞成功,收集、浓缩病毒后测定其滴度为6.7×10~7TU/mL,PCR证实Bdnf基因插入病毒基因组。感染rMSCs后RT-PCR、免疫细胞化学染色及Western检测各组细胞均有BDNF蛋白表达,其中试验组BDNF-rMSCs更大量表达BDNF,与其余2组(Mock-rMSCs、rMSCs)比较差异具有统计学意义。构建带有Bdnf基因慢病毒载体并在大鼠骨髓间质干细胞中成功表达,为今后基因修饰干细胞的移植后长期观察研究奠定了基础。     

大鼠Bdnf基因慢病毒载体的构建及其在骨髓间质干细胞中的表达

骨髓间质干细胞（MSCs）是目前基因工程正在探讨应用的靶细胞,为构建带有脑源性神经营养因子（Bdnf）基因慢病毒载体并使其在大鼠骨髓间质干细胞中表达,采用RT-PCR技术获得大鼠Bdnf基因编码区（CDS）片段,限制性内切酶酶切和基因重组构建慢病毒载体质粒PNL-BDNF-IRES₂-EGFP,在脂质体介导下与包装质粒HELPER,包膜质粒VSVG共转染293T细胞包装生产慢病毒。所获慢病毒感染大鼠MSCs（rMSCs）后,PCR和免疫细胞化学法检测在rMSCs中Bdnf基因的插入和表达。结果显示所获的Bdnf基因经测序后与GenBank报道序列完全一致。重组慢病毒载体质粒PNL-BDNF-IRES2-EGFP经鉴定正确。三质粒共转染293T细胞成功,收集、浓缩病毒后测定其滴度为6.7×10⁷TU/mL, PCR证实Bdnf基因插入病毒基因组。感染rMSCs后RT-PCR、免疫细胞化学染色及Western检测各组细胞均有BDNF蛋白表达,其中试验组BDNF-rMSCs更大量表达BDNF,与其余2组(Mock-rMSCs、rMSCs)比较差异具有统计学意义。构建带有Bdnf基因慢病毒载体并在大鼠骨髓间质干细胞中成功表达,为今后基因修饰干细胞的移植后长期观察研究奠定了基础。     

Isolation of a cDNA encoding a 70 kDa heat-shock cognate protein expressed in vegetative tissues ofArabidopsis thaliana

A cDNA encoding a 70 kDa heat shock cognate protein (hsc70) was isolated fromArabidopsis thaliana by using a rat hsc70 cDNA as probe. Sequence analysis demonstrated the conservation of functional domains and important amino acid residues among hsc70s in plants and animals. The expression of this gene was stress-inducible, and was found at a substantial level during normal growth in root, stem, leaf and flower tissues, but not in siliques. Multiple copies of this gene exist in theArabidopsis genome.     

Chemical Constituents from Fruits of Ligustrum lucidum

Seven compounds were isolated and identified from the fruits of Ligustrum lucidum Ait. Compounds Ⅰ to Ⅵ were isolated for the first time from the plants of this genus. Compounds Ⅶ is a novel compound named nuezhenidic acid whose chemical structure was elucidated by means of spectral analysis and chemical reaction.     

猪流行性腹泻病毒(PEDV)与抗病毒天然免疫

猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)是引起猪流行性腹泻病等肠道疾病的一种动物冠状病毒.PEDV与宿主系统相互作用,特别是其对宿主抗病毒天然免疫调节作用和机制是目前动物冠状病毒研究的基础科学问题之一.基于作者近几年来对人类重要冠状病毒对宿主抗病毒天然免疫系统调节作用的研究,本文对PEDV基因组与编码蛋白主要功能以及PEDV调节宿主抗病毒天然免疫反应及其可能机制的进展和现状进行了分析.与人类冠状病毒相似,PEDV编码的木瓜样蛋白酶(papain like protease,PLP)是一个多功能蛋白酶,除了蛋白酶活性外,还具有去泛素化酶(DUB)活性和宿主干扰素拮抗活性,是PEDV编码的一种新型病毒来源DUB和宿主干扰素拮抗蛋白.这些研究为阐明PEDV对宿主抗病毒天然免疫反应调节作用和其致病机制提供了重要的理论依据,为研制新型PEDV免疫防治措施提供了重要理论基础.     

The disappearance of an hsc70 species in mung bean seed during germination: purification and characterization of the protein

We have purified a 73 kDa protein from the cytosolic fraction of mung bean seeds. It comprises 0.5–1% of the total protein in seeds. This purified protein is a bona fide hsc70 on the basis of several lines of evidence. First, antibodies against bovine brain hsc70 cross-react with the purified 73 kDa protein. Second, the purified protein comigrates on two-dimensional gels with one of the heat-inducible hsc70s in mung bean seedlings. Third, similar to other hsc70 species, the purified 73 kDa protein has a high affinity for ATP. Finally, the hydrolysis of ATP by the purified protein can be stimulated by peptides; ATPase activity increases from 40 nmol/h to 165 nmol/h per mg of protein. The purified mung bean hsc70 autophosphorylates at a substoichiometric level. Moreover, the amount of this hsc70 species diminishes while new species of hsc70s appear after germination, suggesting that the expressionof hsc70 in mung bean is subject to developmental regulation.     

羊草+大针茅草原退化群落优势种群空间点格局分析

草原退化的特征主要表现为群落生产力的大幅下降和植物个体的小型化, 同时, 退化的草原生态系统与一定强度的放牧压力保持平衡而相对稳定。该文应用摄影定位法测定了羊草+大针茅草原退化群落中4个优势种群羊草(Leymus chinensis)、米氏冰草(Agropyron michnoi)、大针茅(Stipa grandis)、糙隐子草(Cleistogenes squarrosa)的空间格局。点格局分析结果表明: 在空间分布格局上, 4个优势种群均偏离完全随机模型和泊松聚块模型, 而符合嵌套双聚块模型, 也就是说, 其空间格局表现为聚集分布, 且在大聚块中分布着较高密度的小聚块。严重退化的草原群落中优势种群的嵌套双聚块空间分布格局当属一种集体行为, 是种群适应过度放牧压力的一种表现形式, 为过度放牧导致的退化草原群落的主要特征之一。这种生态学现象同植物个体小型化一样, 是种群易化(正相互作用)的结果。在过度放牧的胁迫下, 种群通过改变个体性状及个体在空间的分布状况实现自我帮助, 以抵御外界的放牧压力达到自我保护, 从而维持退化草原生态系统与放牧压力间的相对平衡。     

降水变化对内蒙古典型草原地上生物量的影响

降水格局的变化是气候变化影响干旱和半干旱区草原植物群落结构和功能的关键过程。作为植物群落结构和功能的基本组成单位——植物功能群——对气候变化的响应差异明显。有关不同植物功能群如何响应降水格局的研究, 有助于揭示气候变化对干旱与半干旱区生态系统关键功能与过程稳定性的变异机制。该文利用1982-2015年的典型草原群落地上生物量长期动态监测数据及其所对应的气候数据, 系统揭示降水变化对干旱与半干旱区生态系统生产力的影响。主要研究结果如下: 1) 1982-2015年内蒙古典型草原区降水变化特征明显, 主要表现在降水集中度呈显著的降低趋势; 小降水事件(≤5 mm)明显增多。2)降水变化导致群落生物量呈下降趋势。其中, 多年生丛生禾草的生物量呈上升趋势; 一二年生植物、多年生杂类草、多年生根茎禾类草的生物量均呈下降趋势。3)群落生物量与生长季降水量、降水集中度呈显著的正相关关系。各功能群中, 多年生杂类草、半灌木和生长季降水量呈显著正相关关系, 灌木与降水集中度呈显著负相关关系。4)群落生物量与各等级降水的频率相关性均不显著, 但与I-II类(0.1-10 mm)降水贡献率显著负相关。各功能群中, 多年生杂类草与I类降水(0.1-5.0 mm)的发生频率和降水贡献率均呈显著的负相关关系, 与VI类降水(20-25 mm)的发生频率和贡献率均呈显著的正相关关系。多年生根茎禾类草和VIII类降水(>35 mm)的发生频率和贡献率均呈显著的正相关关系。说明小降水事件的增加将显著降低群落的地上生物量, 小降水事件对干旱、半干旱区植被生长具有重要的生态学意义。     

Blue Light and Abscisic Acid Independently Induce Heterophyllous Switch in Marsilea quadrifolia

In natural habitats Marsilea quadrifolia L. produces different types of leaves above and below the water level. In aseptic cultures growth conditions can be manipulated so that leaves of the submerged type are produced continuously. Under such conditions the application of either blue light or an optimal concentration of abscisic acid (ABA) induced the development of aerial-type leaves. When fluridone, an inhibitor of ABA biosynthesis, was added to the culture medium it did not prevent blue light induction of aerial leaf development. During blue light treatment the endogenous ABA level in M. quadrifolia leaves remained unchanged. However, after the plants were transferred to an enriched medium, the ABA level gradually increased, corresponding to a transition in development from the submerged type of leaves to aerial leaves. These results indicate that the blue light signal is not mediated by ABA. Therefore, in the regulation of heterophyllous determination, discrete pathways exist in response to environmental signals.