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Experimental treatment of the laws of heterogeneous catalysis with immobilized yeast cells (Saccharomyces cerevisiae) 固定化酵母细胞多相催化规律的实验处理
Pub Date : 2000-03-01 DOI: 10.1016/S0307-4412(99)00130-2
Peter Grunwald

A series of simple, low-cost experiments is described in this paper that allows students to be introduced to some basic kinetic laws relating to heterogeneous catalysis. Immobilized yeast cells are used as the example and therefore simultaneously offer the opportunity to acquaint the students with the theoretical and practical background of an important branch of biotechnology.

本文介绍了一系列简单、低成本的实验,使学生了解与多相催化有关的一些基本动力学定律。本课程以固定化酵母细胞为例,同时为学生提供了解生物技术这一重要分支的理论和实践背景的机会。
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引用次数: 1
A guided experimental approach to practical molecular pharmacology teaching: demonstration of sequence selectivity of DNA-binding drugs by arrested polymerase chain reaction 分子药理学实践教学的实验指导方法:用阻滞聚合酶链反应演示dna结合药物的序列选择性
Pub Date : 2000-03-01 DOI: 10.1016/S0307-4412(99)00083-7
Roberto Gambari

Here, we report a simple experimental approach for demonstrating sequence-selectivity of DNA-binding drugs by using the polymerase-chain reaction (PCR). The experiments described could be easily and reproducibly performed by medical and science students and biochemistry teachers, do not require complex or expensive instruments and 32P labelled probes or primers. The procedure described could represent a novel approach to practical biochemistry teaching in the field of biochemical pharma-cology.

在这里,我们报告了一种简单的实验方法,通过聚合酶链反应(PCR)来证明dna结合药物的序列选择性。所描述的实验可以由医学和理科学生和生物化学教师轻松重复地进行,不需要复杂或昂贵的仪器和32P标记的探针或引物。所描述的程序可以代表生化药理学领域生物化学实践教学的一种新方法。
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引用次数: 1
A single protein research integrated advanced biochemistry laboratory course; spectroscopic determination of tyrosyl side chain pKa 结合高等生物化学实验课程的单蛋白研究酪氨酸侧链pKa的光谱测定
Pub Date : 2000-03-01 DOI: 10.1016/S0307-4412(99)00127-2
Brian N. Blanchette, Bal Ram Singh

The unique bio-analytical properties of the amino acid tyrosine (Tyr) are the focus of this experiment from the research oriented biochemistry laboratory course at our university. In the present study pKa1, pKa2, and pKa3 values for free Tyr were estimated to be 2.30, 9.40, and 9.97, respectively, when free Tyr was titrated with 1 mM NaOH and 1 mM HCl using a pH meter. Spectrophotometric analysis of the phenolic side chain pKa3 revealed a value of 10.14, which was consistent with the pKas estimated from the pH meter. The results from this experiment will allow students to compare the free Tyr properties with those present in a protein.

本实验以我校研究型生物化学实验课程中氨基酸酪氨酸(Tyr)独特的生物分析特性为重点。在本研究中,当使用pH计用1mm NaOH和1mm HCl滴定游离Tyr时,pKa1、pKa2和pKa3的值分别为2.30、9.40和9.97。对酚侧链pKa3的分光光度分析显示pKa3的值为10.14,与pH计测得的pKa3值一致。这个实验的结果将允许学生比较游离酪氨酸与蛋白质中存在的特性。
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引用次数: 0
A guided experimental approach to practical molecular pharmacology teaching: demonstration of sequence selectivity of DNA-binding drugs by arrested polymerase chain reaction. 分子药理学实践教学的实验指导方法:用阻滞聚合酶链反应演示dna结合药物的序列选择性。
Pub Date : 2000-03-01
Gambari

Here, we report a simple experimental approach for demonstrating sequence-selectivity of DNA-binding drugs by using the polymerase-chain reaction (PCR). The experiments described could be easily and reproducibly performed by medical and science students and biochemistry teachers, do not require complex or expensive instruments and 32P labelled probes or primers. The procedure described could represent a novel approach to practical biochemistry teaching in the field of biochemical pharma-cology.

在这里,我们报告了一种简单的实验方法,通过聚合酶链反应(PCR)来证明dna结合药物的序列选择性。所描述的实验可以由医学和理科学生和生物化学教师轻松重复地进行,不需要复杂或昂贵的仪器和32P标记的探针或引物。所描述的程序可以代表生化药理学领域生物化学实践教学的一种新方法。
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引用次数: 0
Strategies for building criticism skills in undergraduate biochemists 生物化学本科生批评技巧的培养策略
Pub Date : 2000-03-01 DOI: 10.1016/S0307-4412(99)00138-7
Gareth Denyer

This paper describes an easy method for empowering biochemistry students to think critically about research data. The technique encourages students to formulate their own opinions by removing the usually dominant ‘expert’ commentary. The exercise can be done individually or in very large groups, requires no specialist materials and confirmation that articulation and criticism skills have been learnt can be assessed under standard examination conditions. The skills learnt are not discipline specific and, as well as learning how best to read research papers, students also learn something about the research process.

本文描述了一种简单的方法,使生物化学学生能够批判性地思考研究数据。这种方法鼓励学生去掉通常占主导地位的“专家”评论,形成自己的观点。这个练习可以单独进行,也可以在非常大的小组中进行,不需要专业材料,并且可以在标准考试条件下评估已经学习的表达和批评技能。所学到的技能不是特定学科的,除了学习如何最好地阅读研究论文外,学生还学习了一些关于研究过程的知识。
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引用次数: 3
Journal club as a supplement to the undergraduate biochemistry laboratory 期刊俱乐部作为本科生生物化学实验室的补充
Pub Date : 2000-03-01 DOI: 10.1016/S0307-4412(99)00135-1
Mona L. Hall, Adele J. Wolfson

After purification of lysozyme, our biochemistry students write a research proposal that outlines a strategy for studying this enzyme after alteration by site-directed mutagenesis. Despite a literature search that yielded a wealth of background information, students were often overwhelmed by the assignment because they were not familiar with advanced techniques of protein analysis. We therefore developed a series of journal clubs in which teams of students present methods and data found in papers dealing with lysozyme. The five topics for journal clubs include; substrate binding and mechanism; spectroscopic techniques; stability analysis; two-dimensional NMR; and X-ray crystallography. After the adoption of the group talks, the quality of the research proposals improved immensely and students found the assignment to be an educationally rewarding exercise.

在溶菌酶纯化后,我们的生物化学学生写一份研究计划,概述了通过位点定向诱变改变后研究这种酶的策略。尽管进行了文献检索,获得了丰富的背景信息,但由于学生不熟悉先进的蛋白质分析技术,他们常常被作业压垮。因此,我们发展了一系列的期刊俱乐部,在这些俱乐部中,学生团队介绍在处理溶菌酶的论文中发现的方法和数据。期刊俱乐部的五个主题包括:底物结合及机制;光谱技术;稳定性分析;二维核磁共振;和x射线晶体学。采用小组讨论后,研究建议的质量大大提高,学生们发现这项任务是一项有教育意义的练习。
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引用次数: 0
The excellence of turnip mitochondrial fraction. 萝卜线粒体分数的优良性。
Pub Date : 2000-03-01
Vicente, Madeira

Several mitochondrial fractions were screened for suitability in practical experiments designed for students, with the following issues in mind: avoiding the use of animals; minimal expenditure of labour and time; high enzyme activities; accessible instruments and low-cost materials. Turnips and potato tubers were identified as the best materials from which to extract purified mitochondria according to these criteria, with high respiratory activities and integrity maintained during five consecutive days. Mitochondrial respiration was assayed for succinate, exogenous NADH, malate/pyruvate and alpha-ketoglutarate oxidations with excellent results. At the fifth day, the respiratory control was still about 3, the integrity of the outer mitochondrial membrane maintained at values higher than 80%, and the enzymes retained more than 55% of the initial activities.

在为学生设计的实际实验中,筛选了几种线粒体组分的适用性,考虑到以下问题:避免使用动物;花费最少的人力和时间;酶活性高;易于使用的仪器和低成本的材料。根据这些标准,芜菁和马铃薯块茎被确定为提取纯化线粒体的最佳材料,连续5天保持高呼吸活性和完整性。线粒体呼吸测定琥珀酸、外源性NADH、苹果酸/丙酮酸和α -酮戊二酸氧化,结果良好。第5天,呼吸控制仍在3左右,线粒体外膜完整性维持在80%以上,酶活性保持在初始水平的55%以上。
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引用次数: 0
The excellence of turnip mitochondrial fraction. 萝卜线粒体分数的优良性。
Pub Date : 2000-03-01 DOI: 10.1111/J.1539-3429.2000.TB00034.X
Vicente, Madeira
Several mitochondrial fractions were screened for suitability in practical experiments designed for students, with the following issues in mind: avoiding the use of animals; minimal expenditure of labour and time; high enzyme activities; accessible instruments and low-cost materials. Turnips and potato tubers were identified as the best materials from which to extract purified mitochondria according to these criteria, with high respiratory activities and integrity maintained during five consecutive days. Mitochondrial respiration was assayed for succinate, exogenous NADH, malate/pyruvate and alpha-ketoglutarate oxidations with excellent results. At the fifth day, the respiratory control was still about 3, the integrity of the outer mitochondrial membrane maintained at values higher than 80%, and the enzymes retained more than 55% of the initial activities.
在为学生设计的实际实验中,筛选了几种线粒体组分的适用性,考虑到以下问题:避免使用动物;花费最少的人力和时间;酶活性高;易于使用的仪器和低成本的材料。根据这些标准,芜菁和马铃薯块茎被确定为提取纯化线粒体的最佳材料,连续5天保持高呼吸活性和完整性。线粒体呼吸测定琥珀酸、外源性NADH、苹果酸/丙酮酸和α -酮戊二酸氧化,结果良好。第5天,呼吸控制仍在3左右,线粒体外膜完整性维持在80%以上,酶活性保持在初始水平的55%以上。
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引用次数: 4
How do proteins fold? 蛋白质是如何折叠的?
Pub Date : 2000-03-01 DOI: 10.1016/S0307-4412(99)00144-2
Christopher A Smith

This article emphasizes the importance of getting students to understand the ways in which polypeptides fold to form protein molecules with complex higher-ordered structures. Modern views on how this folding occurs in vitro and in the cell are summarized and set within an appropriate biological context.

本文强调让学生了解多肽折叠形成具有复杂高阶结构的蛋白质分子的方式的重要性。关于这种折叠如何在体外和细胞中发生的现代观点进行了总结,并在适当的生物学背景下进行了设置。
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引用次数: 0
A single protein research integrated advanced biochemistry laboratory course; spectroscopic determination of tyrosyl side chain pKa. 结合高等生物化学实验课程的单蛋白研究酪氨酸侧链pKa的光谱测定。
Pub Date : 2000-03-01
Blanchette, Singh

The unique bio-analytical properties of the amino acid tyrosine (Tyr) are the focus of this experiment from the research oriented biochemistry laboratory course at our university. In the present study pK(a(1)), pK(a(2)), and pK(a(3)) values for free Tyr were estimated to be 2.30, 9.40, and 9.97, respectively, when free Tyr was titrated with 1mM NaOH and 1mM HCl using a pH meter. Spectrophotometric analysis of the phenolic side chain pK(a(3)) revealed a value of 10.14, which was consistent with the pK(a)s estimated from the pH meter. The results from this experiment will allow students to compare the free Tyr properties with those present in a protein.

本实验以我校研究型生物化学实验课程中氨基酸酪氨酸(Tyr)独特的生物分析特性为重点。在本研究中,用pH计用1mM NaOH和1mM HCl滴定游离Tyr时,pK(a(1))、pK(a(2))和pK(a(3))值分别为2.30、9.40和9.97。酚侧链pK(a(3))的光度分析结果为10.14,与pH计测得的pK(a)s一致。这个实验的结果将允许学生比较游离酪氨酸与蛋白质中存在的特性。
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