Genetic Engineering

Teacher

Debanjana Saha

Category

Core Courses

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Course Name : Genetic Engineering

Code(Credit) : ABC01(3-1-1)

Course Objectives

  • To strengthen the knowledge on various cloning and expression vectors
  • To impart the importance of vectors in genetic engineering experiments
  • To strengthen the knowledge on various Strategies of gene cloning

Learning Outcomes

  • Understand and explain the concept of the course of genetic engineering including the techniques, applications and limitations.
  • Demonstrate the ability to design recombinant molecules and apply information extracted from various sources to solve the problems.

 

Course Syllabus

Module I:Introduction to Cloning(4hrs)

Overview of Cloning, Purification and Separation of Nucleic Acids – cutting and joining DNA and vectors. Plasmid vectors, phage vectors,cosmids .

Practice: Restriction enzyme digestion of plasmid DNA or lambda DNA

Module II: Genetic manipulation(4hrs)

Manipulation of DNA–Restriction and Modification enzymes, Design of linkers and adaptors.

Practice:Gel purification of RE digested DNA

Module III: Cloning Vectors and their application(4hrs)

Characteristics of cloning and expression vectors based on plasmid and bacteriophage, Vectors for yeast, insect and mammalian systems, Prokaryotic and eukaryotic expression host systems

Practice: Ligation of DNA fragments with cloning vector pUC18 or pBR322.

Module IV: DNA sequencing(5hrs)

Maxxam-Gilbert- Sanger methods. Automated DNA sequencing. PCR technology - concept- types- primer design- analysis of products and applications. DNA-finger printing, SYBR green assay, Taqman assay, Molecular beacons.

Practice:Preparation of competent cells and Transformation into E.coli with recombinant vector.

Module V: cDNA arrays and Micro array technology(4 hrs)

Strategies for the production of recombinant proteins - insulin- human growth hormone- industrially important proteins. Construction of genomic library- cDNA library

Practice:Isolation of recombinants and confirmation of insert DNA in vector.

Module VI:Analysis and manipulation of Gene expression(4hrs)

Analysis of gene expression, analyzing transcription and translation, Analysis of gene function, site Directed Mutagenesis,Transposon Mutagenesis

Practice:Primer design for PCR and amplification of DNA by PCR.

Module VII: Cloning Application(5hrs)

applications – vaccines – human and genetic diseases – transgenics, Small double stranded RNAs; siRNA technology; Micro RNA; Construction of siRNA vectors; Principle and application of gene silencing.

Practice:Southern Hybridisation

 

Session Plan

Session 1

Overview of Cloning

cloning

cloning

Session 2

Purification and Separation of Nucleic Acids
isolation and purification

Session 3

Session 4  (Project)

Analyzing DNA and cloning

Session 5

Plasmid vectors, phage vectors,cosmids

vectors

types of vectors

Session 6(Practice)

Restriction enzyme digestion of plasmid DNA or lambda DNA

Restriction digestion 

Session 7

Genetic manipulation of DNA

genetic manipulation

adapters and linkers

Session 8( Project)

The principles of genetic engineering

Session 9

Restriction and Modification enzymes

restriction enzyme

restrictionendonucleases

Session 10

Design of linkers and adaptors

linker and adapter

adapters and linkers

Session 11( Practice)

Gel purification of RE digested DNA

Restriction Digest Analysis

Session 12

Session 13

Vectors for Yeast and bacteria

BAC ,YAC

types of vectors

Session 14 (Project)

Different types of cloning

Session 15

Bacteriophage vectors

cosmid

types of vectors

Session 16

Session 17(Project)

Whole-genome sequencing

Session 18

Prokaryotic expression host systems
e.coli

expression host system

Session 19

Eukaryotic expression host system

Yeast

expression host system

Session 20(Practice)

Preparation of competent cells and Transformation into E.coli with recombinant vector

preparation of competent cell
cell transformation

Session 21

Maxam gilbert sequencing method

maxam gilbert sequencing

Dna sequencing

Session 22

Session 23

PCR technology - concept- types- primer design- analysis of products and applications.

PCR

Primer Designing

typesofpcr

Session 24(Project)

Impact of cloning on medicine

 

Session 25

Session 26

SYBR green assay,Taqman assay

SYBR green
Taqman assay

Session 27

Molecular beacons.

Molecular becons

Session 28(Practice )

Ligation of DNA fragments with cloning vector pUC18 or pBR322.

ligation of DNA 

Session 29

Strategies for the production of recombinant proteins

recombinant protein

Recombinant proteins

Session 30(Project)

Studies on DNA-modified organisms

Session 31

insulin- human growth hormone- industrially important proteins

insulin production
growth hormone

Recombinant proteins

Session 32

Session 33(Practice)

Isolation of recombinants and confirmation of insert DNA in vector.

isolation of recombinants 

Session 34

gene expression analysis

expression analysis

microarray technology

Session 35( Project)

Gene-editing technologies, explaining their impact on genetic engineering

Session 36

analyzing transcription and translation

gene regulation 

Session 37

Session 38

Transposon Mutagenesis

transposon 

Session 39(Project)

Cloning: positive or negative outcome for future generations?

Session 40(Practice)

. Primer design for PCR and amplification of DNA by PCRPCR design

transposon

Session41

cloning application

cloning application 

Session 42

Session 43(Project)

Human Genome Project.

Session 44

small interfering RNA and micro RNA

Si and mi RNA

RNA interference

Session 45

gene silencing application

gene silencing 

Session46(Practice)

Southern Blotting

Southern blotting 

Case Studies

Case Studies

Case Studies

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