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为了考查营养条件对质粒DNA的生产影响,采用不同的培养基培养含pcDNA33-HBS质粒的大肠杆菌JM109。实验结果表明:碳源、氮源对质粒DNA产量有明显的影响。葡萄糖是质粒合成过程中较佳的碳源,蛋白胨是较佳的氮源。在M9P培养基中,选择合适的硫酸铵浓度对质粒DNA的生产有一定的作用。由于Gly,Asp,Glu能提供合成核苷酸的氮源,在M9G培养基中添加12g/L的Asp,1.0g/L的Glu和0.4g/L的Gly后,经20h培养,质粒产量可达25mg/L。外源核苷也影响质粒DNA的产量,通过添加0.4g/L胞苷与胸苷的混合物(摩尔比为1∶1)到M9P培养基中,质粒DNA的产量可达35mg/L。 相似文献
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一种简单实用的适于大、小质粒的DNA提取方法 总被引:11,自引:0,他引:11
以Birabolm和Doly的质粒提取方法为基础,用醋酸铵取代醋酸钠,沉淀染色体DNA和RNA,用异丙醇取代乙醇沉淀质粒DNA,降低了质粒DNA标本中蛋白质和RNA的含量,减少了标本体积。并发现在质粒DNA标本中加入冷异丙醇或冷乙醇后,在-70℃、-20℃、-10℃、4℃和室温下作用,对质粒DNA的回收量无明显影响,在质粒DNA标本中加入冷异丙醇后,在4℃作用0、10、20和30分钟,对质粒DNA的回收量无明显影响。用该方法提取的质粒DNA可直接用于限制性内切酶消化。用该方法提取大肠埃希氏菌 相似文献
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介绍了一种成本低、步骤少、简单易行的质粒纯化制检工艺。该工艺选择优势产生超螺旋质粒的大肠杆菌菌株以无蛋白质培养基进行发酵罐培养,采用碱裂解法,对质粒制备过程中所用的层析吸附材料、核酸结合溶液、去除内毒素等杂质的方法和浓缩等步骤进行了实用性改进,并建立了相应的检定方法,所得质粒的纯度达到临床级要求。 相似文献
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对格登发现动物细胞全能性的过程进行介绍。20世纪60年代非洲爪蟾细胞全能性的发现和克隆成功为哺乳动物(如绵羊多利等)的克隆奠定了坚实的基础。英国发育生物学家格登爵士最早实现了这些实验,从而推动了动物细胞重编程领域的研究,此外他还研究了基因表达的分子生物学机制和重编程的分子机制。他是20世纪一位伟大的科学大师。 相似文献
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营养条件对pcDNA3\|HBS质粒DNA生产的影响 总被引:7,自引:0,他引:7
为了考查营养条件对质粒DNA的生产影响,采用不同的培养基培养含pcDNA33-HBS质粒的大肠杆菌JM109。实验结果表明:碳源、氮源质粒DNA产量有明显的影响。葡萄糖是质粒合成过程中较佳的碳源,蛋白胨是较佳的氮源。在MP9P培养基中,选择适合的硫酸铵浓度对质粒DNAR的生产有一定作用。由于Gly,Asp,Glu能提供合成核苷酸的氮源,在M9G培养基中添加1.2g/L的Asp,1.0g/L的Glu和0.4g/L的Gly后,经20h培养,质粒产量可达25mg/L。外源核苷也影响质粒DNA的产量,通过添加0.4g/L胞革与胸苷的混合物(摩尔比为1:1)到M9P培养基中,质粒DNA的产量可达35mg/L。 相似文献
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In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980. 相似文献
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N. P. Vesselkin Yu. V. Natochin 《Journal of Evolutionary Biochemistry and Physiology》2010,46(6):592-603
Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms.
The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal
mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization
followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The
mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction
of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms
are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning
of physiological systems and organs of the living organism 相似文献
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