GST-Smad4融合蛋白的表达与纯化

旨在原核表达Smad4基因,纯化获得GST-Smad4融合蛋白。以人表皮HaCaT细胞的cDNA为模板,利用PCR扩增含有BamH I和SalI酶切位点的Smad4基因;然后将其克隆到pGEX-4T-1原核表达载体中,将正确的重组载体转入大肠杆菌BL21(DE3);用IPTG诱导表达,再利用MagneGST particles亲和纯化GST-Smad4融合蛋白;最后通过Western blot鉴定此融合蛋白。结果显示,成功构建pGEX-4T-1-Smad4原核表达载体;30℃条件下,0.2 mmol/L的IPTG能诱导出大量的可溶性GST-Smad4蛋白;经MagneGST particles纯化的GST-Smad4蛋白可被Smad4的抗体特异识别。纯化的GST-Smad4蛋白可用于后续的生物学研究。     

猪激素敏感脂肪酶基因外显子1的遗传多态性分析

采用PCR-SSCP方法检测猪激素敏感脂肪酶（hormone sensitive lipase,HSL）基因外显子1的多态性,并分析其与初生重、断奶重、6月龄重和背膘厚的关联性。根据猪HSL基因的DNA序列（AJ000483）设计5对引物,结果在P5引物对扩增的片段上发现了多态性,并对纯合子进行测序,发现外显子1的874bp处存在G-A转换,且存在3种基因型（AA、AB、BB）。统计结果表明,3种基因型在各品种中的分布不一致,长白猪和大白猪与莱芜猪和沂蒙黑猪比较差异极显著（P〈0．01）,长白猪与大白猪比较,莱芜猪与沂蒙黑猪比较差异均不显著（P〉0．05）。固定效应模型分析结果表明,背膘厚基因型间差异显著（P〈0．05）,而初生重、断奶重和6月龄重基因型间差异均不显著（P〉0．05）。最小二乘分析结果表明,BB基因型个体同AB和AA基因型个体比较背膘厚的差异显著（P〈0．05）,3种基因型在背膘厚的大小排列顺序为BB〈AB〈AA。因此,推测HSL基因对个体胴体瘦肉率存在一定的影响将HSL某因应用于猪育种过程中的标记辅助选择可以加快猪的育种进程。     

Development of polymorphic microsatellite markers in Camellia chekiangoleosa (Theaceae) using 454-ESTs

? Premise of the study: A set of microsatellite markers for Camellia chekiangoleosa was developed and characterized using 454 sequencing technology to study the population genetic structure and the diversity of germplasm collections. ? Methods and Results: Eighteen polymorphic microsatellite markers were identified and tested in 150 individuals from three natural populations of C. chekiangoleosa. Alleles numbered from two to seven, and the observed and expected heterozygosities ranged from 0.100 to 0.760 and 0.133 to 0.809, respectively. ? Conclusions: These markers will potentially be conducive to further genetic studies on C. chekiangoleosa.     

Habenula "cholinergic" neurons co-release glutamate and acetylcholine and activate postsynaptic neurons via distinct transmission modes

Acetylcholine is an important neurotransmitter, and the habenulo-interpeduncular projection is a major cholinergic pathway in the brain. To study the physiological properties of cholinergic transmission in the interpeduncular nucleus (IPN), we used a transgenic mouse line in which the light-gated cation channel ChannelRhodopsin-2 is selectively expressed in cholinergic neurons. Cholinergic axonal terminals were activated by light pulses, and postsynaptic responses were recorded from IPN neurons. Surprisingly, brief photostimulation produces fast excitatory postsynaptic currents that are mediated by ionotropic glutamate receptors, suggesting wired transmission of glutamate. By contrast, tetanic photostimulation generates slow inward currents that are largely mediated by nicotinic acetylcholine receptors, suggesting volume transmission of acetylcholine. Finally, vesicular transporters for glutamate and acetylcholine are coexpressed on the same axonal terminals in the IPN. These results strongly suggest that adult brain "cholinergic" neurons can corelease glutamate and acetylcholine, but these two neurotransmitters activate postsynaptic neurons via different transmission modes.     

异丙肾上腺素和氨甲酰胆碱对心肌延迟后除极的影响

用乙酰毒毛旋花子甙元2.0×10~(-7)mol/L,在绵羊心室浦肯野纤维产生延迟后除极(DAD)为实验模型,观察异丙肾上腺素与氨甲酰胆碱对DAD的影响。异丙肾上腺素1.0—3.0×10~(-8)mol/L能增加DAD幅值,为剂量依赖性,并可诱发出触发型心律失常。氨甲酰胆碱2.0×10~(-6)mol/L单独作用对DAD幅值无影响,然而在异丙肾上腺素增强DAD幅值或产生触发型心律失常后,同样浓度的氨甲酰胆碱能显著地减小DAD幅值以及消除触发型心律失常。但氨甲酰胆碱对由高钙、氨茶碱和组胺等所增加的DAD幅值却无抑制作用。以上结果表明,氨甲酰胆碱能拮抗异丙肾上腺素引起的DAD增强作用,可能与M受体的激活,并继而降低膜上β受体激活时所提高的腺苷酸环化酶活性有关。     

四川白水河自然保护区问题分析排序与对策研究

以白水河国家级自然保护区为研究对象,采用“问题树模型”、“对策树模型”和“距阵排序”等分析工具,对白水河自然保护区过去存在的问题进行分析与评价,并提出了白水河自然保护区的具体保护管理对策。     

叶绿体——疫苗生产新工厂

以叶绿体为受体的叶绿体基因工程技术已经在工农业及医药生物技术领域发挥了重要作用。利用叶绿体为基因转移受体的叶绿体基因工程技术来生产疫苗是目前人们关注的焦点。现着重在叶绿体作为遗传工程受体的特点、叶绿体基因工程的研究进展,它在疫苗生产中的应用及发展前景等方面进行综述。     

醋酸钙复盐对微生物生长的影响

应用四醋酸钙对黑曲霉、黄曲霉、米曲霉、链格孢霉等霉菌和大肠杆菌、枯草杆菌、绿脓杆菌、金黄色葡萄球菌等细菌生长影响作了研究,并将四醋酸钙与醋酸钙、苯甲酸钠对上述微生物的生长抑制作用加以比较。0.4％四醋酸钙对上述细菌的生长有明显的抑制作用。对上述霉菌生长亦有一定的抑制作用。当其与苯甲酸钠协同使用时,能更明显地抑制微生物生长。实验结果表明四醋酸钙很可能是一种安全性高、成本低的新型食品防腐剂。     

The influence of water quality and sediment geochemistry on the horizontal and vertical distribution of phosphorus and nitrogen in sediments of a large,shallow lake

Distinct horizontal water column concentration gradients of nutrients and chlorophyll a (Chl a) occur within large, shallow, eutrophic Lake Taihu, China. Concentrations are high in the north, where some of the major polluted tributaries enter the lake, and relatively low in the south, where macrophytes generally are abundant. It is not clear, however, whether these water column concentration gradients are similarly reflected in spatial heterogeneity of nutrient concentrations within the bottom sediments. The main objective of this study was therefore to test if horizontal and vertical variations in the phosphorus and nitrogen content in bottom sediments of Lake Taihu are significantly related to (1) horizontal variations in overlying water column nutrient concentrations and (2) other sediment geochemical constituents. We measured the concentration of total phosphorus (TP) and total nitrogen (TN) in surficial sediments (0–2 cm) and TP, TN and Chl a concentrations in water column samples, collected from 32 sites in 2005. In 2006 sediment, TP, TN, carbon, iron and manganese concentrations were measured vertically at 2 cm intervals, extending to a depth of approximately 20 cm, at an additional eight sites. Linear correlation analysis revealed that surficial sediment TP concentrations across the 32 stations were related significantly, though weakly, to annual mean water column concentrations of TP, TN as well as Chl a. Correlations of surficial sediment TN with water column variables were, however, not significant (P > 0.05). Amongst the geochemical variables tested, the vertical variability of sediment TP concentrations was most strongly related to sediment manganese and carbon concentrations. A multiple stepwise linear regression revealed that the combination of sediment manganese and carbon concentrations explained 91% of the horizontal variability in sediment TP concentrations and 65% of the vertical variability. Handling editor: Luigi Naselli-Flores