单核细胞增多性李斯特菌的脉冲场凝胶电泳分型研究

用限制性内切酶Sma Ⅰ和Apa Ⅰ分别对21株单核细胞增多性李斯特菌菌株基因组DNA进行酶切,用脉冲场凝胶电泳（PFGE）方法进行遗传多样性分析。结果表明：3株临床分离株（00181、00174和0194）分别存在于亚型A、B、D中;16个单核细胞增多性李斯特菌奶相关和环境分离株主要存在于4个亚型中,其中2个亚型（C、D）均为奶相关分离株（包括原料奶、均质奶（储奶罐）和成品奶分离株）,而加工厂地面污水分离株为独立的一个亚型（A）。     

Conformational dynamics of individual DNA molecules during gel electrophoresis

Gel electrophoresis is widely used in molecular biology to separate DNA molecules according to their sizes. The physical basis of this size separation is, however, poorly understood. Here we report observations of individual, fluorescently stained DNA molecules as they migrate during various kinds of gel electrophoresis. Their movement, under the influence of either a steady electric field or a pulsed-field, is characterized by cycles of elongation and contraction. Initially relaxed coils of DNA lengthen into 'hook-shaped' configurations which temporarily 'hang-up' on obstacles in the gel matrix before sliding off, contracting and entering another cycle. The effects of a new electrophoresis technique, termed 'pulse-oriented electrophoresis', which allows the effective angle of the electric field, and hence the molecular orientation of DNA, to be varied without electrode rearrangement, are also studied. In this case the DNA adopts a 'staircase' configuration showing that the net orientation in a direction is given by the vector sum of the pulses used.     

聚丙烯酰胺凝胶电泳体系中蛋白质的荧光标记分析

在室温下,采用聚丙烯酰胺凝胶电泳法,分离经荧光胺或对氯荧光胺荧光标记的蛋白质样品,可在2.5小时内完成蛋白质样品的分离和其区带的鉴定。耗时是传统方法的四分之一,而且荧光图象清晰、灵敏度较高。     

聚丙烯酰胺凝胶电泳快速显色方法的研究

通过对聚丙烯酰胺凝胶电泳的固定方法、时间、染色液种类、染色方法及蛋白灵敏度的研究,确定了以10%三氯乙酸为固定液,室温固定10min的固定方法;染色液为：考马斯亮蓝G-250、95%乙醇和10%磷酸,染色时间为15min;蛋白质灵敏度与考马斯亮蓝R-250相当,达到0.1μg。     

十二烷基硫酸钠-聚丙烯酰胺凝胶电泳的实验教学改进

在本科生物化学实验教学中,为减少有机溶剂的使用,同时缩短实验时间,作者对SDS-PAGE的脱色过程进行改进,使用0.5mol/L NaC l代替含挥发性有机溶剂做脱色液,取得了良好的实验效果。     

Abnormal pattern detected in fragile-X patients by pulsed-field gel electrophoresis.

The fragile-X syndrome is the most frequent inherited form of mental retardation, with an incidence of 1 in 1,500 males. It is characterized by the presence of a fragile site at Xq27.3 induced in vitro by folate deprivation or by inhibitors of deoxynucleotide synthesis. Its mode of inheritance is unusual for an X-linked trait, with incomplete penetrance in both males and females. Some phenotypically normal males transmit the mutation to all their daughters who rarely express any symptoms, but penetrance is high in sons and daughters of these carrier women. Genetic and physical mapping of the Xq27-q28 region has confirmed that the disease locus is located at or very near the fragile site. Hypotheses proposed to account for the abnormalities in the inheritance of the disease include sequence rearrangements by meiotic recombination or a mutation that affects reactivation of an inactive X chromosome during differentiation of female germ cells. To detect such rearrangements, or methylation changes that may reflect a locally inactive X chromosome, we used pulsed-field gel analysis of DNA from fragile-X patients with probes close to the fragile-X locus. The probe Do33 (DXS465) detected abnormal patterns in fragile-X patients, but not in normal controls or in non-expressing male transmitters.     

改良聚丙烯酰胺凝胶电泳法测定籼粳分化的同工酶带

对同工酶聚丙烯酰胺凝胶电泳法进行改进:液氮研磨样品,0.1 mol/L tris-顺丁烯二酸酐(pH=7.0)缓冲液提取,Bio-rad MP3电泳槽(凝胶规格10 cru×7.5 cm×0.5 mm)在120～150 V电泳2～4 h,得到快速、经济、分辨率高的分析效果.分析了水稻4种同工酶,发现Est-3,Est-4,Est-x和Pox-x4个新的籼粳分化位点,其中Pox-x为一新位点,该位点在籼稻中有1条带,在粳稻中为哑等位基因.