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1 Hereditas (Beijing) 2016 年 5 月, 38(5): 遗传学教学人类血型性状综合遗传大实验的设计与教学实践赵健 1, 胡冬梅 1, 于大德 2, 董明亮 1, 李云 1, 范英明 1, 王延伟 1, 张金凤摘要 : 综合大实验能够训练学生灵活应用理论知识并掌握实验技能, 成为当前实验课教学改革的重要方式本文以人类的 ABO 血型性状为实验对象, 设计了人类 ABO 血型分子基因分型与群体遗传平衡分析大实验实验中提取同学唾液中黏膜细胞的 DNA, 经过 PCR 扩增目的片段酶切及电泳分离一系列分子遗传技术分析, 鉴定出每位同学的基因型 ; 然后以全班同学为一个类似孟德尔群体调查 ABO 血型的各种基因型频数, 用 Popgene 软件分析各种群体遗传参数通过开放教学不仅让学生掌握了分子遗传实验技术和群体遗传分析技术及软件应用, 还让学生自主设计方案优化分子技术环节, 提高学生驾驭知识的能力通过 5 年的教学探索与实践, 建立了稳定的分子遗传实验体系, 能够清楚地检测出 ABO 血型的 6 种基因型 :I A I A I A i I B I B I B i I A I B ii; 综合了分子遗传与群体遗传的实验教学, 统计全班同学 6 种基因型的频数, 直接计算 3 个复等位基因的频率, 进而应用软件分析群体遗传各种参数 ; 实现了学生自主设计并完成实验的开放式实验教学 ; 大实验教学获得了学生的好评, 取得了很好的教学效果该大实验可直接应用于生物类专业的遗传学实验教学, 其中的教学理念和方法还可推广应用于其他生物学实验教学关键词 : 综合大实验 ;ABO 血型 ; 自主设计 ; 开放式实验教学 Teaching design and practice of human blood type traits in genetics comprehensive laboratory course Jian Zhao 1, Dongmei Hu 1, Dade Yu 2, Mingliang Dong 1, Yun Li 1, Yingming Fan 1, Yanwei Wang 1, Jinfeng Zhang 1 1. College of Biological Sciences and Technology, Beijing Forestry University, Beijing , China; 2. Department of Forest Botany and Tree Physiology, Georg-August-University Göttingen, Göttingen 37077, Germany Abstract: Comprehensive laboratory courses, which enable students to aptly apply theoretic knowledge and master experiment skills, play an important role in the present educational reform of laboratory courses. We utilized hu- 收稿日期 : ; 修回日期 : 基金项目 : 2015 ( YWKC15013)( 2015ZCQ- SW-02) ( J ) ( bjfubio2014) [Supported by the 2015 Beijing Forestry University Graduate Course Development Project (No.YWKC15013) the Medium and Long Scientific Research Project for Young Teachers in Beijing Forestry University(No. 2015ZCQ-SW-02) the National Natural Science Foundation of China (No. J ) and the Education Reform Project of Beijing Forestry University (No. bjfubio2014)] 作者简介 : zhaojian0703@hotmail.com 通讯作者 : zjf@bjfu.edu.cn DOI: /j.yczz 网络出版时间 : 2016/1/12 15:58:11 URL:

2 462 Hereditas (Beijing) man ABO blood type as the experimental subject, and designed the experiment Molecular Genotyping of Human ABO Blood Type and Analysis of Population Genetic Equilibrium. In the experiment, DNA in mucosal cells is extracted from students saliva, and each student s genotype is identified using a series of molecular genetics technologies, including PCR amplification of target fragments, enzymatic digestion, and electrophoretic separation. Then, taking the whole class as an analogous Mendel population, a survey of genotype frequency of ABO blood type is conducted, followed with analyses of various population genetic parameters using Popgene. Through the open laboratory course, students can not only master molecular genetic experimental skills, but also improve their understanding of theoretic knowledge through independent design and optimization of molecular techniques. After five years of research and practice, a stable experimental system of molecular genetics has been established to identify six genotypes of ABO blood types, namely I A I A, I A i, I B I B, I B i, I A I B and ii. Laboratory courses of molecular and population genetics have been integrated by calculating the frequencies of the six genotypes and three multiple alleles and testing population genetic equilibrium. The goal of the open laboratory course with independent design and implementation by the students has been achieved. This laboratory course has proved effective and received good reviews from the students. It could be applied as a genetics laboratory course for the biology majors directly, and its ideas and methods could be promoted and applied to other biological laboratory courses. Keywords: comprehensive experiment; ABO blood type; independent design; open laboratory course [1] [2] [3] [4] [5] ABO

3 5 : 463 ABO ABO 4 A B AB O A B I A I B O i A I A I A I A i B I B I B I B i ABO [6] 1990 ABO [7] ABO 6 I A I A I A i I B I B I B i I A I B ii ( ) ABO DNA [8] PCR DNA [9] 3 3 PCR 2~3 Hardy-Weinberg Popgene ~5 1 DNA PCR DNA DNA PCR DNA

4 464 Hereditas (Beijing) 图 1 以人类血型性状综合遗传大实验为例的分子遗传和群体遗传的教改流程图 Fig. 1 Flow chart of course design for molecular genetics and population genetics educational reform (in the case of comprehensive genetic experiments of human blood type traits) PCR Popgene ( 1) 3 DNA DNA 100 ng/μl PCR DNA 1 DNA ng/μl ng/μl DNA 2 PCR 159 bp 200 bp 2% PCR 200 bp Kpn 159 bp Alu DNA Kpn 200 bp Alu 159 bp A Kpn 200 bp Alu 118 bp B Kpn 171 bp Alu 159 bp i ZTZX DNA ABO PCR 54 3% 5 11 ZTZX 3% PCR ABO Popgene

5 5 : A B C ABO PCR PCR DNA DNA DNA 4 5 (1) ABO 6 I A I A I A i I B I B I B i I A I B ii (2) 6 3 (3) (4) (5) [10] [2,3] ABO

6 466 Hereditas (Beijing) ABO 附录 : 参考文献 (References): [1] The Department of Education s opinions on comprehensively improving higher education quality. The Department of Education (2012) No [2012] [2] Polanyi M. The Study of Man. London: Routledge & Kegan Paul, 1957: 12. [3] Zhang YY. Theoretical basis exploration and analysis of practical teaching. China Univ Teach, 2006(6): 彥.., 2006(6): [4] Shi W. On the university practical teaching system. J High Educ, 2013, 34(7): , 2013, 34(7): [5] Zhang JF, Hu DM, Zheng CX, Zhao N. Education practice and exploration of genetics opening laboratory course// Anthology of University Teaching Forum on Life Science. Beijing: Beijing Forestry University, 2009: ,,,. //. :, 2009: [6] Liang QJ, Wang C. Analysis of genetic equilibrium of ABO blood type. Bull Biol, 2008, 43(1): ,. ABO., 2008, 43(1): [7] Yamamoto F. ABO blood group system ABH oligosaccharide antigens, anti-a and anti-b, A and B glycosyltransferases, and ABO genes. Immunohematology, 2004, 20(1): [8] Chen JC, Zhang HY, Peng JY, Fu YY, Ye WC, Chang H. Simple method and application for DNA extraction from saliva. J Sun Yat-sen Univ (Med Sci), 2006, 27(4S): ,,,,,. DNA. ( ), 2006, 27(4S): [9] Liu CL, Miao TH, Gong XY, Wang ZY, Ren FR, Lv QS, Liu JH. Application of multiplex PCR on ABO genotyping. Beijing Med J, 2007, 29(2): ,,,,,,. PCR ABO., 2007, 29(2): [10] The State Council General Office s implementation opinions on deepening enterprising education reform in institutions of higher learning. The State Council General Office (2015) No. 36, [2015]36, ( 责任编委 : 陈德富 )

7 5 : 467 附图 1 Fig. S1 实验进程考核单 Checklist of experimental process 附图 2 生科 12 班两名同学的 DNA 提取检测结果 Fig. S2 Testing results of DNA extraction of two students in Class 12

8 468 附图 3 Fig. S3 Hereditas (Beijing) 2016 第 38 卷生科 12 班 8 名同学的 2 对引物 PCR 片段电泳图 The electrophoretogram of PCR fragments amplified by two pairs of primers of eight students in Class 12 附图 4 生科 12 班 4 名同学的酶切片段电泳结果及这 4 名同学的血型基因型鉴定结果 Fig. S4 The electrophoretogram results of digestion fragments of 4 students in Class 12 and their genotypic identification results of blood types 左侧: 1-4 表示学生编号 200 bp 等是 DNA 片段长度右侧: 1-4 号学生血型基因型鉴定结果

9 5 : 469 附图 5 生科 11 班 ZTZX 小组不同退火温度 PCR 扩增产物电泳结果 Fig. S5 Electrophoresis result of PCR amplification products with different annealing temperature of Group ZTZX in Class 11 附表 1 Popgene 软件的生科 12 班 ABO 血型基因型的群体遗传参数分析 Table S1 Population genetic parameter analysis of ABO blood type in Class 12 by Popgene A 为基因型统计结果及卡方值计算 ;B 为遗传平衡检验结果 ;C 为等位基因频率 A Population: 1 Locus: p1 Genotypes Obs. (O) Exp. (E) (O-E)2/E 2*O*Ln(O/E) (A, A) (B, A) (B, B) (i, A) (i, B) (i, i) B C Allele Frequency of population 1 Chi-suqare for Hardy-Weinberg equilibrium Allele\ Locus p1 Chi-square: Allele A Degree of freedom: 3 Allele B Probability: Allele i Likelihood ratio test for Hardy-Weinberg equibrium:: G-square: Degree of freedom: 3 Probability:

Microsoft Word - 793-797 tb20150504赵宏宇s-高校教改纵横.doc

Microsoft Word - 793-797 tb20150504赵宏宇s-高校教改纵横.doc 微生物学通报 Microbiology China tongbao@im.ac.cn Apr. 20, 2016, 43(4): 793 797 http://journals.im.ac.cn/wswxtbcn DOI: 10.13344/j.microbiol.china.150504 高校教改纵横生物工程专业发酵课程群建设探索 * 赵宏宇