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Course Detail

Course Name Advanced Molecular Genetics and Gene Expression Lab
Course Code 25BIO481
Program B.Sc. (Hons.) Microbiology
Semester 7
Credits 2
Campus Amritapuri

Syllabus

  1. Culturing and Handling of Drosophila:  
    1. Media Preparation  
    2. Cleaning and Sterilization of bottles  
    3. Handling of Drosophila
  2. Morphology and Sexual dimorphism.    
  3. Salivary gland Chromosome- Dissection of Salivary glands and Preparation of Polytene chromosome.  
  4. Problems on:  
    1. Sex linkage  
    2. Genetic map  
  5. Karyotyping (Optional) 
  6. Heterologous expression of recombinant proteins and purification in coli
    1. PCR amplification of target gene and cloning in inducible vectors 
    2. Chemical mediated transformation/electroporation 
    3. Purification of recombinant protein by affinity chromatography 
  7. Analysis of fluorescent labelled bacteria  
  8. Transformation of Sacharomyces cerevisae by electroporation and selection of recombinants 

Objectives and Outcomes

LEARNING OBJECTIVES

 

  1. Understand the principles and methods for culturing and genetically analyzing Drosophila melanogaster as a model organism.
  2. Gain practical knowledge of genetic techniques such as sex linkage analysis, genetic mapping, and chromosome visualization.
  3. Learn the fundamentals of molecular cloning, protein expression, and purification in bacterial and yeast systems.
  4. Acquire hands-on skills in transformation, fluorescence analysis, and recombinant protein handling.

COURSE OUTCOMES:

CO1: Maintain and handle Drosophila cultures, and differentiate between sexes based on morphological traits.

CO2: Perform salivary gland dissection and prepare polytene chromosome slides for cytogenetic analysis.

CO3: Solve classical genetics problems including sex linkage and genetic mapping using Drosophila.

CO4: Clone a target gene into an inducible vector, transform E. coli, and express and purify the recombinant protein using affinity chromatography.

CO5: Analyze fluorescent-labeled bacterial cells and transform Saccharomyces cerevisiae, selecting recombinants through appropriate screening techniques.

Text Books / References

REFERENCE:

  1. Green, M. R., & Sambrook, J. (2012). Molecular Cloning: A Laboratory Manual (4th ed.). Cold Spring Harbor Laboratory Press.

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