DNA recovery from agarose gels with a simple centrifuge-driven sephadex filtration

Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and effective recovery of DNA from agarose gels with good yield and quality. Using a Sephadex resin filled spin column, DNA fragments of 500 bp to 6 kb in an agarose gel slice were easily recovered by a 2 min centrifugation. The recovery efficiencies were over 40%-50% and the eluted DNA can be used directly for downstream application, such as polymerase chain reactions (PCR) and restriction enzyme digestion. This method could also be used to recover large DNA fragment (48 kb) without degradation. The use of Sephadex helps to remove small molecular impurities from agarose and it also reduces the chance of clogging the column filter caused by direct contact with agarose.     

干旱胁迫对白桦实生幼苗叶片蛋白质的影响

The studies on leave protein of Betula platyphylla seedlings by the methods of SDS-PAGE electrophoresis and dimensional gel electrophoresis was concerned. The result indicated that some new proteins such as 29kD pI4.3, 29kD pI4.2, 14kD pI5.3 and 11.5kD pI5.16 protein point were induced by soil drought stress, and some protein disappeared,for instance four protein between 12.4 - 11.5kD pI5.8 - 6.44 in seedlings leaves of B. platyphylla during soil drought stress.     

双向电泳在植物蛋白质组中的应用及影响因素分析

双向电泳(two-dimensional gel electrophoresis,2-DE)是蛋白质组学研究中常用的技术之一,因具有较高的稳定性和重复性,在植物蛋白质组学中应用广泛。蛋白质双向电泳(2-DE)过程复杂、时间持久以及影响因素众多,本文立足蛋白质双向电泳实验过程中可能出现的各种现象,如：分离的蛋白质点过少、条纹过多导致凝胶背景模糊、高丰度蛋白点聚集沉淀及低丰度蛋白点被掩盖等,着重分析其产生的原因并提出具体的解决办法,为蛋白质双向电泳技术初学者提供参考。     

基于不同回收方法的肌浆蛋白对鲢鱼糜冻融稳定性的影响

为研究不同回收方法的肌浆蛋白对鲢鱼糜冻融稳定性的影响,分别通过加热法、酸偏移法和酸偏移-耦合壳聚糖絮凝法回收鲢鱼糜漂洗液中的肌浆蛋白,并将其添加至鱼糜中,测定鱼糜冻融循环过程中蛋白质冷冻变性、脂肪氧化及凝胶品质的指标。结果显示:添加不同方法回收的肌浆蛋白均能抑制鲢鱼糜冻融循环过程中盐溶性蛋白含量、Ca²⁺-ATPase活性和总巯基含量的下降以及表面疏水性的上升,其中加热法回收的肌浆蛋白抑制鱼糜蛋白质冷冻变性的效果最显著,经过9次冻融后仍能够对鱼糜蛋白质变性起到抑制效果。蛋白质羰基含量测定结果显示,肌浆蛋白的添加可以抑制鱼糜冻融循环过程中蛋白质氧化现象。而与空白组鱼糜相比,添加不同回收处理的肌浆蛋白的鱼糜,其硫代巴比妥酸值（TBARs）和pH值并没有显著变化。此外,随着冻融循环次数的增加,鱼糜凝胶品质会发生严重劣化,其中添加酸偏移-耦合壳聚糖絮凝处理的肌浆蛋白的鱼糜始终保持较好的凝胶质构性能。     

A quick method for the assessment of activity and inhibition of fish amylases

A sensitive and quick method was developed to determine the presence of α-amylase in the gut of aquatic organisms, as well as its sensitivity to inhibitors. The assay is based on the utilization of Petri dishes filled with starch–agarose gel as a substrate for the enzyme solution, which is placed in small wells punched in the surface. Circular zones produced by the action of amylase remain colourless after staining with lugol. Pure commercial porcine amylase was used to fit the better conditions for developing the assay (1 g L^–1 starch in the gels, 4 h of incubation). The diameter of the cleared zones were related to the activity of enzyme and the method detected linearly amylase activity in a range of 2–20 U well^–1, so it was used to reveal the presence of amylase in digestive extracts obtained from different sparid fish. The method was also used to evaluate the effect produced by a specific inhibitor on fish amylases, showing a linear response when the ratio inhibitor:enzyme (in units) changed from 20:1 to 2:1. Comparison of the cleared zones produced by amylases of sparid fish in the presence or absence of inhibitor, revealed differences in their sensitivity to inhibition, which ranged from 15 to 50% of total activity. The assay is proposed for a preliminary evaluation of possible inhibitors contained in feedstuffs used in fish feeding.     

脱氢松香基有机小分子凝胶因子的合成及其性能

用12-溴-13,14-双硝基脱氢松香酸甲酯合成了一种新型的有机小分子凝胶因子1,4a-二甲基-6-溴-1,2,3,4,4a,17,18,18a-八氢萘基[2,1-h]二吡啶并[3,2-a:2,3-c]吩嗪甲酸甲酯(DBNPM),通过1H NMR,13C NMR,MS和IR等表征手段对产物结构进行了表征。为了考察此小分子凝胶因子的自组装行为,研究了该分子在不同有机溶剂中的成胶性能和形貌。研究发现,在甲醇中临界成胶浓度(CGC)最低,为0.5 g/L;但是在二氧六环中凝胶性质最为稳定。扫描电镜(SEM)照片显示,从甲醇溶剂中得到的干胶呈片状结构,从二氧六环溶剂中得到的干胶呈网状结构;通过红外光谱和XRD测试研究证实,该化合物分子间的氢键是凝胶化的重要驱动力之一。