The topics covered in this course will include the following: (i) basics of genetic engineering such as enzymes used in genetic engineering (restriction enzymes, DNA ligases, DNA polymerases, etc.), cloning vectors (plasmid vectors, phage vectors, etc.), genotypes and gene transfer methodologies (transformation and transfection), methodologies of gene cloning (genome cloning, cDNA cloning, PCR, chemical synthesis of DNA, etc.), and analyses of gene (hybridization analyses, blotting analyses, etc.); (ii) applications of genetic engineering such as gene diagnosis and DNA test, gene therapy, metagenome, genetically-modified plants, clone animals, stem cells, and regenerative medicine.
This course has two aims. The first is to teach students the basics of genetic engineering that are required during laboratory work. The second is to introduce the hot topics on applications of genetic engineering, and to realize the contribution of genetic engineering to human welfare.
At the end of this course, students will be able to:
1) Have an understanding of basics of genetic engineering such as restriction enzyme, cloning vector, host, gene cloning, DNA sequencing.
2) Have an understanding of applications of genetic engineering such as gene diagnosis and DNA test, gene therapy, clone animal, stem cell.
Restriction enzyme, Cloning vector, Host, Gene cloning, DNA sequencing, Gene diagnosis and DNA test, Gene therapy, Clone animal, Stem cell.
|✔ Specialist skills||Intercultural skills||Communication skills||Critical thinking skills||✔ Practical and/or problem-solving skills|
Students are required to read the related section of text book before coming to class. Every class will be given by using PowerPoint slides, board writing, handouts, etc. Students are given exercise problems and reports during class as necessary (less than 20%).
|Course schedule||Required learning|
|Class 1||Enzymes used in genetic engineering (I): restriction enzymes and DNA ligases||Explain the reaction and classification of restriction enzymes, and the reaction of DNA ligases.|
|Class 2||Enzymes used in genetic engineering (II): DNA polymerases, etc.||Explain the reaction of DNA polymerases and other enzymes.|
|Class 3||Plasmid vectors (I): plasmid vectors, methodologies of transformation||Explain the structure of plasmid vectors and the methodologies of transformation.|
|Class 4||Plasmid vectors (II): applications of plasmid vectors, preparation of plasmd||Explain how to use plasmid vectors and how to prepare plasmid DNA.|
|Class 5||DNA sequence analysis: DNA sequencing (Maxam and Gilbert method, Sanger method, DNA sequencers, and next-generation DNA ) Midterm examination||Explain the methodologies of DNA sequencing.|
|Class 6||Methodologies of gene cloning: genome cloning, cDNA cloning and cloning of PCR products||Explain the methodologies of genome cloning, cDNA cloning and how to clone PCR products.|
|Class 7||Phage vectors: phage vectors, methodologies of transfection, applications of phage vectors||Explain the structure of phage vectors, the methodologies of transfection, and how to use phage vectors.|
|Class 8||Gene expression and functional analyses of genes (I): optimization of gene expression, expression of fusion genes, and phage display||Explain the optimization of gene expression, expression of fusion genes, and phage display.|
|Class 9||Gene expression and functional analyses of genes (II): analyses of transcription initiation point and DNA binding sequences, and RNAi||Explain the analyses of transcription initiation point and DNA binding sequences, and RNAi.|
|Class 10||Analyses of gene (I): electrophoresis, hybridization analyses (Southern, northern and colony hybridizations)||Explain the hybridization analyses.|
|Class 11||Analyses of gene (II): expressed protein analyses (Western blotting and ELISA)||Explain expressed protein analyses.|
|Class 12||Applications of genetic engineering (I): gene diagnosis and DNA test, gene therapy, and metagenome||Explain the gene diagnosis and DNA test, gene therapy, and metagenome.|
|Class 13||Applications of genetic engineering (II): genetically-modified plants, clone animals, stem cells and regenerative medicine||Explain the genetically-modified plants, clone animals, stem cells and regenerative medicine.|
|Class 14||Applications of genetic engineering (III): genetically-modified plants and regenerative medicine Endterm examination||Explain the genetically-modified plants and regenerative medicine.|
To enhance effective learning, students are encouraged to spend approximately 100 minutes preparing for class and another 100 minutes reviewing class content afterwards (including assignments) for each class.
They should do so by referring to textbooks and other course material.
Hiroshi Nojima, "Genetic Engineering, from Basics to Applications", Tokyo, Tokyo Kagakudojin, ISBN 978-4-8079-0804-2 (Japanese)
Handouts will be distributed at the beginning of class when necessary.
Students will be assessed on their understanding of basics and applications of genetic engineering.
Students’course scores are based on exercise problems, reports and so on.
Students must have successfully completed Biochemistry I (LST.A203), Biochemistry II (LST.A218), Molecular Biology I (LST.A208) and Molecular Biology II (LST.A213), or have equivalent knowledge.
Eiry Kobatake: kobatake.e.aa[at]m.titech.ac.jp, Masayasu Mie: mie.m.aa[at]m.titech.ac.jp
Rie Yatsunami: ryatsuna[at]bio.titech.ac.jp,
Contact by e-mail in advance to schedule an appointment.