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BMS537 Methods In Recombinant DNA Technology UITM Assignment Sample Malaysia

This course at UITM introduces students to the fundamental toolbox of recombinant DNA technology. Through a combination of hands-on lab sessions, lectures, and tutorials, students will gain practical experience and understanding of various techniques, including DNA extraction, gel electrophoresis, gene cloning, restriction enzyme mapping, DNA hybridization, DNA sequencing, and polymerase chain reaction. Emphasis is placed on explaining the mechanisms behind these techniques.

The hands-on approach in workshop-style lab sessions ensures that students develop essential skills and knowledge to explore advanced techniques and comprehend the applications of recombinant DNA technology in research. By the end of the course, students will be equipped with the necessary expertise to employ these methods effectively and engage in further studies or research in the field.

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Assignment Brief 1 : Explain the mechanisms underlying basic methods in recombinant DNA technology

Recombinant DNA technology, also known as genetic engineering or gene cloning, involves manipulating DNA molecules to create new combinations of genetic material. It has revolutionized various fields, including biotechnology, medicine, and agriculture. The fundamental mechanisms underlying basic methods in recombinant DNA technology are:

  • Restriction Enzymes: These are specialized proteins that can cut DNA at specific recognition sequences. They act as molecular scissors, cleaving DNA molecules into fragments with sticky ends (single-stranded overhangs) or blunt ends. Restriction enzymes play a crucial role in DNA cloning and gene manipulation.
  • DNA Ligation: DNA fragments created by restriction enzymes can be reconnected using DNA ligases. Ligases catalyze the formation of phosphodiester bonds between the sugar and phosphate groups of adjacent DNA fragments, resulting in a recombinant DNA molecule.
  • Transformation: In genetic engineering, transformation refers to the process of introducing foreign DNA into a host organism. Bacterial cells are commonly used as host organisms because they can take up DNA from their surroundings. The recombinant DNA is introduced into the bacterial cells, which then replicate, propagating the foreign DNA.
  • Polymerase Chain Reaction (PCR): PCR is a technique that allows the rapid amplification of specific DNA sequences. It involves cycles of denaturation, annealing, and extension using DNA polymerases. PCR is invaluable for generating large amounts of DNA for further analysis or cloning.
  • Plasmids and Vectors: Plasmids are small, circular DNA molecules found in bacteria and other microorganisms. We serve as vectors in recombinant DNA technology, carrying foreign DNA into host cells. Plasmids often contain antibiotic resistance genes, which allow researchers to select for cells that have successfully taken up the recombinant DNA.
  • Gel Electrophoresis: This technique separates DNA fragments based on their size and charge. DNA samples are loaded onto a gel matrix and subjected to an electric field, causing the fragments to migrate through the gel. Smaller fragments move faster, enabling researchers to analyze and isolate specific DNA segments. 

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Assignment Brief 2 : Illustrate using specific examples how recombinant DNA technology can be used to solve problems

  • Human Insulin Production: Recombinant DNA technology has enabled the production of human insulin using genetically engineered bacteria. Previously, insulin was extracted from animal sources, leading to potential allergic reactions in patients. By inserting the human insulin gene into bacterial cells, they can now produce and purify large quantities of human insulin, ensuring a safer and more reliable supply for diabetic patients.
  • Golden Rice: Vitamin A deficiency is a significant health issue in developing countries, leading to blindness and other health problems. To address this, scientists used recombinant DNA technology to insert genes responsible for beta-carotene synthesis (a precursor of vitamin A) into rice plants, creating “Golden Rice.” This genetically modified crop helps combat vitamin A deficiency by providing a dietary source of the essential nutrient.
  • Environmental Cleanup: Recombinant DNA technology has been employed to develop microorganisms capable of degrading harmful pollutants in the environment. For instance, oil-eating bacteria were genetically engineered to break down oil spills more effectively, offering a potential solution to environmental disasters caused by oil pollution.
  • Biopharmaceutical Production: Through recombinant DNA technology, human cells or bacteria can be modified to produce therapeutic proteins and antibodies. This approach has revolutionized the pharmaceutical industry by enabling large-scale production of medicines, such as growth hormones, blood-clotting factors, and monoclonal antibodies used to treat various diseases.

Assignment Brief 3 : Perform experiments in basic recombinant DNA techniques CLO4 Demonstrate written communication skills in scientific writing

For this experiment, we will perform a simple cloning procedure using plasmids and Escherichia coli (E. coli) bacteria as the host organism. The goal is to insert a gene of interest into a plasmid and then introduce it into the bacterial cells for replication.

Materials:

  • Plasmid DNA containing a selectable marker (e.g., antibiotic resistance gene)
  • DNA fragment of the gene of interest
  • Restriction enzymes specific to the gene of interest and plasmid
  • DNA ligase
  • E. coli bacterial cells
  • LB agar plates with appropriate antibiotics

Experimental Steps:

Isolation and Purification of Plasmid DNA:

Extract plasmid DNA from a bacterial culture containing the plasmid of interest using a plasmid extraction kit.

Isolation and Purification of Gene of Interest: 

Extract the gene of interest from the source organism’s DNA (e.g., human genomic DNA) using appropriate extraction and purification methods.

Digestion of Plasmid and Gene of Interest:

  •  Incubate the purified plasmid DNA and the gene of interest with specific restriction enzymes that cut at desired recognition sites.
  • The restriction sites on the plasmid and the gene of interest should be compatible for proper ligation.

Ligation of Plasmid and Gene of Interest:

 Mix the digested plasmid and gene of interest together and add DNA ligase to catalyze the ligation of their sticky ends or blunt ends.

Transformation of E. coli Cells:

  • Heat shock E. coli competent cells with the ligated DNA to facilitate the uptake of the recombinant plasmid by the bacterial cells. 
  • Plate the transformed bacterial cells on LB agar plates containing an appropriate antibiotic for the selectable marker present in the plasmid.

Selection and Verification: 

  • Incubate the plates overnight at the appropriate temperature for bacterial growth.
  • The transformed cells will grow into colonies, and only those containing the recombinant plasmid with the gene of interest will survive on the antibiotic-containing plates. 
  • Pick a few colonies, perform plasmid isolation, and confirm the presence of the gene of interest using PCR or other suitable methods.

This experiment demonstrates the fundamental steps of recombinant DNA technology, from DNA manipulation using restriction enzymes and DNA ligase to the transformation and selection of genetically modified bacterial cells. It showcases how this technology can be used to create genetically modified organisms with desired traits.

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