Course title
遺伝子工学   [Gene Engineering]
Course category		Requirement		Credit	2
Department		Year	3～	Semester	1st
Course type	1st	Course code	01BN3127
Instructor(s)
松下 保彦, 佐々木 信光   [MATSUSHITA Yasuhiko, SASAKI Nobumitsu]
Facility affiliation	Center for Infectious Disease Epidemiology and Prevention Research	Office	Gene Res. Center East Bld.: room E17 and E18	Email address

Course description

This course describes basic techniques relating to a recombinant DNA experiment, analysis of gene expression, and functional analysis of gene products.

Expected Learning

Learners who successfully complete this course will be able to: (1) Recognize the basic techniques for recombinant DNA experiments. (2) Make experimental plans for the research using recombinant DNA technologies including cloning and functional analysis of genes. (3) Understand the principle of representative experimental method used for analysis of gene expression and functional analysis of gene products. Corresponding criteria in the Diploma policy: See the curriculum maps. https://www.tuat.ac.jp/campuslife_career/campuslife/policy/

Course schedule

========================== Weeks 1 to 7 [by Sasaki] ========================== 1. Gene cloning method (1) ・ Conventional cloning method using restriction enzymes and modification enzymes ・ Latest cloning method by in vitro recombination method ・ Frequency and efficiency of transformation of E. coli 2. Gene cloning method (2) ・ Analysis of DNA base sequence (Sanger method [dideoxy method]) ・ Introduction and utilization of tag sequences ・ Planning of gene cloning experiment 3. Gene amplification method (1) ・ PCR ・ cDNA synthesis by reverse transcription reaction ・ Primer design ・ Mutagenesis using PCR (1) 4. Gene amplification method (2) ・ Mutagenesis using PCR (2) ・ RACE method ・ Inverse PCR method 5. Library construction ・ Genomic library ・ cDNA library ・ Equalized cDNA library ・ Full length cDNA library ・ Subtraction method, Differential display method ・ Colony & Plaque Hybridization 6. Analysis method of gene expression (1) ・ Northern blot analysis ・ Western blot analysis, ELISA method 7. Analysis method of gene expression (2) ・ Real time quantitative PCR method ・ Digital PCR method ================================ Weeks 8 to 15 [by Matsushita] ================================ 8. Analysis method of gene expression (3) ・ Overview of the analysis methods of nucleic acids ・ Outline of microarray experiment ・ DNA chip (GeneChip) ・ Gene expression analysis using DNA microarray 9. Analysis method of gene expression (4) ・ Principle of next generation DNA sequencer ・ Gene expression analysis by next generation DNA sequencer 10. Analysis method of gene expression (5) ・ Analysis of chromatin immunoprecipitation with next generation DNA sequencer 11. Analysis method of gene expression (6) ・ Overview of the analysis methods of proteins ・ Gene expression analysis by mass spectrometer (protein) 12. Functional Analysis Method of Gene Products (1) ・ Overview of functional analysis methods of genes ・ Points of functional analysis of genes 13. Functional Analysis Method of Gene Products (2) ・ Knockout method and knockdown method (gene disruption, tag-line, RNAi) ・ Enhancer trap method ・ Use of artificial mutant protein (Partial deletion type, dominant negative type modification protein) ・ Gene expression system in E. coli (Protein Expression System and Its Utilization, Principle of Affinity Purification, Vector system for inducible expression) ・ Cell-free translation system ・ Gene introduction method Physical and chemical methods (gene gun, electroporation, drug treatment, Freeze-thaw) Biological methods (binary vector method, viral vector method) 14. Functional Analysis Method of Gene Products (3) ・ Analysis of protein-nucleic acid interaction (Yeast one hybrid method, South western method, Chromatin immunoprecipitation analysis, Footprint method Electrophoresis mobility shift assay [= EMSA method]) 15. Functional Analysis Method of Gene Products (4) ・ Analysis method of protein-protein interaction (Yeast two hybrid method, Far Western method, FRET method, BiFC method, immunoprecipitation method)

Prerequisites

It is preferable to have taken molecular biology I and II. In addition to 30 hours in the class, students are recommended to prepare for and revise the classes spending the standard amount of time as specified by the University.

Required Text(s) and Materials

By E-file acquisition from moodle or print distribution.

References

Molecular Biology of the Gene (ISBN: 978-0-321-76243-6) Principles of genetic engineering (ISBN: 978-4-7827-0637-4) (Sankyo Publishing)

Assessment/Grading

Evaluate by examination (100%). Two faculty members in charge will conduct the examination in each of the responsible times.

Message from instructor(s)

If you are interested, please study actively by reading reference books more and more.You are recommended to read the original text of reference books (in English), to get used to English if possible.

Course keywords

Cloning, gene amplification, gene introduction, vector, library, next generation genome analyzer, analysis of gene expression, function analysis of gene products

Office hours

Taking an appointment by e-mail or after class. Sasaki: chaki@cc.tuat.ac.jp, Matsushita: ymatsu@cc.tuat.ac.jp

Remarks 1

Remarks 2

Distribution of past results are as follows. H30 (2018) S 34%, A 38%, B 16%, C 8%, D 0%, E 4% H29 (2017) S 38%, A 28%, B 18%, C 6%, D 4%, E 6% H28 (2016) S 24%, A 28%, B 18%, C 20%, D 8%, E 1% H27 (2015) S 29%, A 12%, B 41%, C 17%, D 0%, E 1% H26 (2014) S 23%, A 31%, B 25%, C 16%, D 3%, E 2% H25 (2013) S 19%, A 19%, B 25%, C 19%, D 8%, E 10% H24 (2012) S 16%, A 17%, B 14%, C 24%, D 15%, E 14% H23 (2011) S 9%, A 20%, B 19%, C 39%, D 13% H22 (2010) S 12%, A 17%, B 27%, C 23%, D 21% H21 (2009) S 6%, A 42%, B 33%, C 4%, D 15% H20 (2008) S 3%, A 32%, B 50%, C 11%, D 4% H19 (2007) S 15%, A 24%, B 30%, C 20%, D 11% H18 (2006) S 27%, A 33%, B 21%, C 13%, D 6% H17 (2005) S 9%, A 26%, B 32%, C 21%, D 12%

Related URL

Lecture Language

Japanese

Language Subject

Last update

3/5/2019 6:06:47 PM