| Course title | |||||
| 遺伝子工学 [Gene Engineering] | |||||
| Course category | Requirement | Credit | 2 | ||
| Department | Year | 3～ | Semester | Spring | |
| Course type | Spring | 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 Research Center, East building, 2nd Floor | Email address | |
| Course description |
| This course describes basic techniques relating to a recombinant DNA experiment and outlines how the individual technique is being used in each application of medical treatments and industries. |
| 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) Develop the viewpoint to understand how genetic engineering techniques are applied to medical treatments and industries, from application or research examples. |
| Course schedule |
|
========================== Weeks 1 to 6 [by Sasaki] ========================== ＜Gene amplification methods＞ ・PCR ・cDNA synthesis by reverse transcription ・primer design ・mutation introduction method using PCR ＜Gene cloning methods＞ ・conventional cloning method using restriction enzyme and modification enzyme ・the latest cloning method by in vitro recombination method ・frequency and efficiency of transformation of E. coli ・conventional DNA sequencing method (Sanger method [dideoxy method]) ・addition and utilization of tag sequences ・experimental planning of gene cloning in E. coli ＜library＞ ・enome library, cDNA library ・Equalized cDNA library, full length cDNA library ・subtraction method, differential display method ・colony＆plaque hybridization, selection by antibody ＜Cloning method of adjacent unknown sequence fragment using known sequence＞ ・RACE ・inverse PCR ・walking PCR ＜Gene expression assays＞ ・northern and western blot analyses ・enzyme-linked immunosorbent (ELISA) assay ・quantitative real-time RT-PCR ・digital PCR method ================================ Weeks 7 to 13 [by Matsushita] ================================ < Analysis of gene expression > ・Outline of the microarray experiment ・DNA chip (GeneChip) ・Gene expression analysis using DNA microarray ・The principle of next-generation DNA sequencer ・The gene expression analysis by next-generation DNA sequencer ・The chromatin immunoprecipitation analysis using next-generation DNA sequencer < Function analysis of the gene product > ・knockdown and knockout methods (gene disruption, tag line, RNAi) ・Enhancer trap method ・use of artificial mutant protein (partial deletion type, dominant negative form of protein modification) ・Gene expression system in E. coli (protein production system, the principle of affinity purification, inducible expression vector system) ・Cell-free translation system ・Method of gene transfer; physical and chemical methods (gene gun, electroporation, chemical treatment, freeze-thaw) biological method (binary vector method, virus vector method ) ・Analysis of the interaction of proteins and nucleic acids (yeast one-hybrid method , south western blots, chromatin immunoprecipitation analysis, footprinting electrophoresis mobility shift assay [= EMSA method]) ・Analysis of protein-protein interactions (yeast two-hybrid method, far western method, FRET method,BiFC method, immunoprecipitation method) ========================== Week 14 Examination ========================== |
| Prerequisites |
| It is preferable to have taken molecular biology I and II. |
| 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) |
| Assessment/Grading |
| By the evaluation of (20%) attendance and (80%) examination. |
| 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 |
| 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 |
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Distribution of past results are as follows. 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 |
| 3/22/2017 5:57:36 PM |