B.Sc. Microbiology Semester 5: Genetic Engineering and rDNA Technology

BT5005: Genetic Engineering and rDNA Technology

• The course aims to impart fundamental knowledge about theoretical aspects and applications of rDNA technology and genetic engineering for human benefits.

Objectives

• Acquire adequate knowledge and necessary skills related to different techniques involved in genetic engineering.
• Understand the objective of gene cloning, library preparation, and screening of positive clones from the library.
• Learn about advanced techniques of Recombinant DNA Technology and genetic engineering.

Course Outcomes

• CO1: Understand the players of gene cloning, including DNA modifying enzymes, cloning vectors, and expression vectors.
• CO2: Learn the steps of gene cloning, library preparation, and screening of positive clones from the library.
• CO3: Understand advanced techniques of Recombinant DNA Technology, including blotting, DNA sequencing, PCR, and mutagenesis.
• CO4: Acquire knowledge about recombinant proteins, gene therapy, and advanced techniques of genetic engineering.

Course Structure

• Unit I: Introduction to Gene Cloning (12 hours)
  • Molecular tools for gene cloning, including enzymes, nucleases, and ligases.
  • Cloning vectors and their desirable properties.
  • Expression vectors, including pET, pMAl, and pGEX.
• Unit II: Strategies of Genetic Engineering (12 hours)
  • Isolation of DNA to be cloned, including genomic and plasmid DNA.
  • Construction of genomic and cDNA libraries.
  • Methods of gene transfer, including transformation and growth of cells.
  • Selection of clones and recombinant selection methods.
• Unit III: Advanced Techniques of rDNA Technology (12 hours)
  • Blotting techniques, including Southern, Northern, and Western blotting.
  • DNA sequencing, PCR, and real-time PCR.
  • Mutagenesis, protein engineering, and protein-DNA interactions.
• Unit IV: Applications of Genetic Engineering (12 hours)
  • Recombinant products, including therapeutic proteins and vaccines.
  • Applications of gene cloning in agriculture, medicine, and environmental and forensic science.
  • Gene therapy.

Practical

• Isolation of genomic DNA and quantification.
• PCR amplification and analysis by agarose gel electrophoresis.
• Isolation of total RNA and quantification.
• Preparation of plasmid, pET-28a, and analysis.
• Restriction digestion analysis of a given plasmid.
• Preparation of competent cells and transformation in E. coli.

Reference Books

• Biotechnology: The Biological Principles by M.D. Trevan et al.
• Biotechnology by Smith, Cambridge Press.
• Advanced Molecular Biology by Twyman R.M.
• Microbiology by Atlas R.M.
• Microbiology- Prescott L.M.
• Microbial Genetics by Freifelder D.
• Principles of Gene Manipulation by Old and Primrose.
• Principles of Gene Manipulation and Genome by Primrose and Twyman.

Programme Outcomes

• PO1: Knowledge of microbiological sciences and their application in industry and hospitals.
• PO2: Core Competence in microbiology, including the ability to find remedies for diseases.
• PO3: Breadth of knowledge in microbiology, including its impact on human health and the environment.
• PO4: Preparation for research projects and industrial applications.
• PO5: Professionalism in microbiology, including career opportunities.
• PO6: Evaluation of academic performance based on continuous internal evaluation and semester-end examinations.

BT5005: Genetic Engineering and rDNA Technology

• The course aims to impart fundamental knowledge about theoretical aspects and applications of rDNA technology and genetic engineering for human benefits.

Objectives

• Acquire adequate knowledge and necessary skills related to different techniques involved in genetic engineering.
• Understand the objective of gene cloning, library preparation, and screening of positive clones from the library.
• Learn about advanced techniques of Recombinant DNA Technology and genetic engineering.

Course Outcomes

• CO1: Understand the players of gene cloning, including DNA modifying enzymes, cloning vectors, and expression vectors.
• CO2: Learn the steps of gene cloning, library preparation, and screening of positive clones from the library.
• CO3: Understand advanced techniques of Recombinant DNA Technology, including blotting, DNA sequencing, PCR, and mutagenesis.
• CO4: Acquire knowledge about recombinant proteins, gene therapy, and advanced techniques of genetic engineering.

Course Structure

• Unit I: Introduction to Gene Cloning (12 hours)
  • Molecular tools for gene cloning, including enzymes, nucleases, and ligases.
  • Cloning vectors and their desirable properties.
  • Expression vectors, including pET, pMAl, and pGEX.
• Unit II: Strategies of Genetic Engineering (12 hours)
  • Isolation of DNA to be cloned, including genomic and plasmid DNA.
  • Construction of genomic and cDNA libraries.
  • Methods of gene transfer, including transformation and growth of cells.
  • Selection of clones and recombinant selection methods.
• Unit III: Advanced Techniques of rDNA Technology (12 hours)
  • Blotting techniques, including Southern, Northern, and Western blotting.
  • DNA sequencing, PCR, and real-time PCR.
  • Mutagenesis, protein engineering, and protein-DNA interactions.
• Unit IV: Applications of Genetic Engineering (12 hours)
  • Recombinant products, including therapeutic proteins and vaccines.
  • Applications of gene cloning in agriculture, medicine, and environmental and forensic science.
  • Gene therapy.

Practical

• Isolation of genomic DNA and quantification.
• PCR amplification and analysis by agarose gel electrophoresis.
• Isolation of total RNA and quantification.
• Preparation of plasmid, pET-28a, and analysis.
• Restriction digestion analysis of a given plasmid.
• Preparation of competent cells and transformation in E. coli.

Reference Books

• Biotechnology: The Biological Principles by M.D. Trevan et al.
• Biotechnology by Smith, Cambridge Press.
• Advanced Molecular Biology by Twyman R.M.
• Microbiology by Atlas R.M.
• Microbiology- Prescott L.M.
• Microbial Genetics by Freifelder D.
• Principles of Gene Manipulation by Old and Primrose.
• Principles of Gene Manipulation and Genome by Primrose and Twyman.

Programme Outcomes

• PO1: Knowledge of microbiological sciences and their application in industry and hospitals.
• PO2: Core Competence in microbiology, including the ability to find remedies for diseases.
• PO3: Breadth of knowledge in microbiology, including its impact on human health and the environment.
• PO4: Preparation for research projects and industrial applications.
• PO5: Professionalism in microbiology, including career opportunities.
• PO6: Evaluation of academic performance based on continuous internal evaluation and semester-end examinations.

This quiz covers the concepts of genetic engineering and rDNA technology for B.Sc. Microbiology students in their 5th semester.