1
课程详述
COURSE SPECIFICATION
联系授课教师。
The course information as follows may be subject to change, either during the session because of unforeseen
circumstances, or following review of the course at the end of the session. Queries about the course should be
directed to the course instructor.
1.
课程名称 Course Title
生物化学与分子生物学实验/Biochemistry and Molecular Biology Laboratory
2.
授课院系
Originating Department
生物系/Department of Biology
3.
课程编号
Course Code
BIO222
4.
课程学分 Credit Value
2
5.
课程类别
Course Type
专业基础课(生物科学、生物技术专业) - Major Foundational Courses (Biological
Sciences, Biotechnology)
专业选修课(生物信息专业) - Major Elective Courses (Bioinformatics)
6.
授课学期
Semester
春季 Spring
7.
授课语言
Teaching Language
英文 English / 中英双语 English & Chinese
8.
他授课教师)
Instructor(s), Affiliation&
Contact
For team teaching, please list
all instructors
赵颖岚 工程师
南方科技大学生物系
广东省深圳市西丽区南方科技大学第一教学楼 229
Tel (电话): (0755)8801-8749
Email: zhaoyl@sustc.edu.cn
Dr. ZHAO Yinglan, Engineer, Department of Biology
Teaching building 1, room 229
马小英 工程师
南方科技大学生物系
广东省深圳市西丽区南方科技大学第一教学楼 231
Tel (电话): (0755) 8801-8746
Email: maxy@sustc.edu.cn
Dr. MA xiaoying, Engineer, Department of Biology
Teaching building 1, room 231
9.
/
方式
Tutor/TA(s), Contact
贾方兴 实验员
南方科技大学生物系
广东省深圳市西丽区南方科技大学第一教学楼 232
Tel (电话): (0755) 8801-0325
Email: Jiafx@sustc.edu.cn
Mr. JIA Fangxing, Laboratory Technician, Department of Biology
2
Teaching building 1, room 232
林飚 实验员
南方科技大学生物系
广东省深圳市西丽区南方科技大学第一教学楼 234
Tel (电话): (0755) 8801-8469
Email: Linb3@sustc.edu.cn
Mr. JIA Fangxing, Laboratory Technician, Department of Biology
Teaching building 1, room 234
10.
选课人数限额(不填)
Maximum Enrolment
Optional
授课方式
Delivery Method
习题/辅导/讨论
Tutorials
实验/实习
Lab/Practical
其它(请具体注明)
OtherPlease specify
总学时
Total
11.
学时数
Credit Hours
课堂实验 61 学时
61 class hours
3
时)
3 class hours observation
64
12.
先修课程、其它学习要求
Pre-requisites or Other
Academic Requirements
要求先修普通生物实验、《生物 I》课;学习本验课同时学习分子
物学》课程。
Pre-study General Biology Laboratory and Biochemistry I, and study the Molecular
Biology with the experiment course at the same time.
13.
后续课程、其它学习规划
Courses for which this course
is a pre-requisite
14.
其它要求修读本课程的学系
Cross-listing Dept.
教学大纲及教学日历 SYLLABUS
15.
教学目标 Course Objectives
蛋白质、糖类、脂类、核酸等生物化学分子是生命体组成成分及各种生理活动的执行者。解析它们的结构、特性和功能是
解读生命的一个重要方面。随着现代科学技术的发展,各种先进的手段愈来愈被广泛地应用于生物化学研究中。生物化学
实验已是生物系各专业的必修实验课。本课程设置具有层次性,每部分内容能相互衔接,使整个生物化学实验课程形成一
个有机的整体。课程内容分为五个模块:生化实验基本技能、核酸模块、蛋白模块,酶动力学模块,碳水化合物模块;内
容涉及蛋白质、酶、糖类、核酸等常见生物大分子的提取、分离、检测、功能和应用等。
Experiments in this lab are continues integrity focused on biomolecules, help the students to grasp modern biochemistry
and molecular biology techniques. Study the target gene from gene level (molecular cloning), protein level (recombinant
protein expression and purification in E. Coli system) and its function (enzyme kinetics analysis). The experiments also
include basic concept of acids, bases and buffers, DNA melting curve, to get the student a comprehensive understanding
of biology in chemical angle.
16.
预达学习成果 Learning Outcomes
给学生提供了一个通过实验了解和探索常见生化大分子结构、特性和功能的平台,锻炼学生定性观察和定量检测等技能;
希望学生通过对本课程的学习,培养动手能力、设计实验的创新思维及分析解释实验现象,初步培养学生的科学的批判精
神和素养。
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The laboratory is chosen to give students an opportunity to explore the structure, characteristic and function of
biomolecules. With the experiments, practise the ability of qualitative observation and quantitative detection.We hope it
can improve students’ practical ability, innovative thinking in experiment design and ability to analyse and solve
problems. At the same time, it can train the students’ critical thinking in scientific work.
17.
课程内容及教学日历 (如授课语言以英文为主,则课程内容介绍可以用英文;如团队教学或模块教学,教学日历须注明
主讲人)
Course Contents (in Parts/Chapters/Sections/Weeks. Please notify name of instructor for course section(s), if
this is a team teaching or module course.)
一、生物化学与分子生物学实验简介及溶液配制(4 学时)
Lab 1. Introduction to Biochemistry and Molecular Biology Laboratory and preparation of solutions (4 credit hours)
介绍实验室规则、安全、课程设置及实验安排;实验记录本及实验报告要求;实验课考核及评分标准;初步介绍整个本课
程核酸模块及蛋白模块实验基础知识。了解生物化学与分子生物学实验中常用仪器的使用;学习溶液配制,了解溶液配制
的计算方法,学生自己计算、称量以及配制实验二所需溶液。
Introduce Biochemistry and Molecular Biology Laboratory safety, course requirement (notebook and report), course
schedule (basic content of nucleic acid and protein) and evaluation. To know the usage of basic equipments will meet in
this class; Learn how to prepare solutions, and know the methods of calculation. The students should calculate, weight
and prepare the solutions needed in experiment 2.
二、酸、碱及缓冲液(5 学时)
Lab 2. Acids, Bases and Buffers (5 credit hours)
学习 pH 计的校准及使用;配制醋酸缓冲液和 Tris-Cl 冲液;了解缓冲液对酸、碱的缓冲能力及稀释对缓冲液缓冲能力的
影响;计算缓冲液的缓冲能力;滴定法计算氨基酸的滴定曲线。
To learn how to standardize and perform pH meter; prepare acetate buffer and tris-HCl buffer with the solutions made in
last class. To study the influence of acid, base and dilution to buffer, and calculate the buffer capacity of buffer. To
understand the buffer region by calculating titration curve of unknown amino acids.
三、测定 DNA 溶解曲线(4 学时)
Lab 3. DNA melting curve (Calf thymus DNA solution) (4 credit hours)
了解 DNA 效应效应 pH 牛胸 DNA 解曲线测不强度小牛 DNA
溶解曲线;比较不同条件对 DNA 溶解曲线的影响及其在生物学中的作用。
To learn hyperchromicity and hypochromicity of DNA, compare the melting curve of calf thymus DNA solution in high pH
and different ionic strength with control to understand the effects of different conditions to DNA melting curve and its
function.
四、质粒提取及限制性内切酶酶切(5 学时)
Lab 4.Plasmid purification and Restriction Enzyme Digestion (5 credit hours)
介绍分子克隆基础识及质粒提取理(着重介绍 123 种成的作用,理解醇沉淀原理等);Nanodrop
定提取质粒 DNA 浓度;对提取质粒进行限制性内切酶酶切,特别强调微量操作方法。
Introduction the basic knowledge of molecular cloning and principle of preparation of plasmid DNA by alkaline lysis with
SDS. To let the student understand the function of each element in solution 1, 2, 3 and the principle of ethanol
precipitation. To detect the DNA concentration with Nanodrop and then digest with plasmid with EcoRI and XhoI. In this
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experiment, we especially focus on submicroliter fluid handling.
五、琼脂糖凝胶电泳及胶回收(5 学时)
Lab 5. Restriction endonuclease digestion, Electrophoresis, Gel extraction (5 credit hours)
了解琼脂糖凝胶电泳的原理;琼脂糖凝胶电泳制备;通过琼脂糖凝胶电泳观察提取的质粒及酶切产物;胶回收试剂盒回收
酶切后目的 DNA 片段。
To understand the principle for agarose gel electrophoresis. To know how to prepare agarose gel and observe the result
of electrophoresis - plasmid and restriction enzyme digest products. Cut the desired fragment from agarose gel and do
gel extraction to get pure DNA fragments.
六、连接及转化(5 学时)
Lab 6. Ligation and Transformation (5 credit hours)
掌握连接及转化的基本原理;将胶回收产物(目的片段及载体)进行连接;连接产物转化大肠杆菌。课堂实验 4 学时,第
二天观察 1 学时。
To master the basic principle for ligation and transformation. Ligate target fragment and vector (extract from agarose gel)
and transformed toE.coli. 4 class hours for the classroom teaching and 1 class hour for the results observed in the next
day.
七、菌落 PCR 筛选阳性克隆及引物设计(5 学时)
Lab 7. Screening PCR and Primer design (5 credit hours)
掌握菌落 PCR 筛选阳性隆的原理;菌 PCR 扩增平板上克;琼脂糖凝胶泳检 PCR ,筛选阳性克;了
引物设计基本原理及应用。
To master the principle for screening positive colonies by the method of colony PCR. Detect the PCR results with
agarose gel electrophoresis to select the positive colonies. To study the principle and application of primer design.
-九、蛋白表达及 SDS-聚丙烯酰胺凝胶电泳(9 学时)
Lab 8&9. Protein expression and SDS-PAGE (9 credit hours)
掌握大肠菌表达外蛋白的原并掌 SDS-聚丙酰胺凝胶泳分离蛋质的原理制样方法在不同条下诱
肠杆菌表达目的蛋白;样品制备。SDS-聚丙烯酰胺凝胶电泳及染色观察,检测蛋白表达。课堂实验 8 学时,第二天观察 1
学时。
To learn the principle of foreign protein expression in E.coli and SDS-PAGE. Induce the protein expression under
different conditions and harvest the cells. Cast SDS-PAGE gel and prepare sample for electrophoresis. To run the SDS-
PAGE and stain the gel, detect the protein expression level. 8 class hours for the classroom teaching and 1 class hour
for the results observed in the next day.
十、His-Tag 融合蛋白纯化(5 学时)
Lab 10. Purify a His-Tag protein and SDS-PAGE (5 credit hours)
学习亲和层析的原理,重点学习 Ni-柱纯化 His-Tag 融合蛋白的原理;Ni 柱纯化融合蛋白;SDS-丙烯酰胺凝胶电泳检
纯化产物。课堂实验 4 学时,第二天观察 1 学时。
To learn the principle of affinity chromatograph, especially Ni-column purify His-tagged fusion protein. Using Ni-column
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purify the target protein and detect the purity by SDS-PAGE. 4 class hours for the classroom teaching and 1 class hour
for the results observed in the next day.
十一、凝胶过滤层析与 Bradford 法定量蛋白质(4 学时)
Lab 11. Gel-filtration chromatography & Protein Quantification by Bradford Assay (4 credit hours)
介绍凝胶过滤层析的原理及其在本实验中的实验目-脱盐纯化;根据聚丙烯酰胺凝胶电泳结果选取纯度及浓度最佳的样本
进行蛋白定量。学习不同蛋白定量原理,着重学习 Bradford 法及紫外分光光度计法。利用标准曲线, excel 中作图,对
目的蛋白定量。
To introduce the principle of gel-filtration chromatography and understand the usage in this experiment -desalting (for
purification); Choose the best sample with highest concentration and purity for protein quantification. To learn the
principle of protein quantification with different methods, especially focus on Bradford and UV-vis spectroscopy assay.
Draw the standard curve in excel and calculate the protein quantification.
十二、乳酸脱氢酶动力学(5 学时)
Lab 12. Kinetics of LDH (5 credit hours)
理解酶动力学特性;运用分光光度计法检测乳酸脱氢酶 KM Vmax;计算乳酸脱氢酶的比活力。
To understand the characteristic of enzyme kinetics. To detect the KMand Vmaxof LDH and calculate enzyme activity.
十三、DNS 法检测还原性糖(4 学时)
Lab 13.Determination of Reducing Sugar with DNS (4 credit hours)
线不同
用、酸水解后还原性单糖的含量。
To master the principle of carbohydrate quantification, and take advantage of standard curve to determine the
concentration of different sugar solution. Compare the reducing sugar content by Hydrolysis with acid and enzyme
digest.
十四、实验回顾与总结(4 学时)
Lab 14. Review (4 credit hours)
学生根据课程内容对实验原理、步骤、实验结果进行回顾,并对本实验相关内容进展扩展,以演讲形式讲解。
The students will summary the lab course by themselves and expand the mind. They will do the presentation with PPT.
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教材及其它参考资料 Textbook and Supplementary Readings
Biochemistry Laboratory: Modern Theory and Techniques, 2nd. (R.F. Boyer, 2011)
Lehninger Principles of Biochemistry, 7th. (D.L. Nelson, M.M. Cox, 2017)
Laboratory Manual of Biochemistry: Methods and Techniques. (R.S. Sengar, 2014)
课程评估 ASSESSMENT