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BMS424 Microbial Molecular Genetics UITM Assignment Answer Malaysia

BMS424 Microbial Molecular Genetics is a course offered at UITM Malaysia that provides students with an introduction to the fundamental molecular aspects of prokaryotic systems. The course delves into the processes of DNA replication, transcription, gene recombination, and protein translation, emphasizing their underlying molecular mechanisms. Additionally, the course covers gene expression and manipulation, offering insights into how genes are controlled and engineered. Students will gain a comprehensive understanding of microbial genetics at the molecular level, enabling them to explore the intricate world of microorganisms and their genetic makeup. 

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Assignment Activity 1 : Explain the structure and organization of nucleic acids, as well as the processes of DNA replication, protein synthesis and gene transfer in microorganism. 

Structure and Organization of Nucleic Acids:

 Nucleic acids are macromolecules essential for storing, transmitting, and expressing genetic information in living organisms. Two main types of nucleic acids exist: deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). DNA is the genetic material responsible for hereditary information, while RNA plays a crucial role in protein synthesis and other cellular processes.

The basic building blocks of nucleic acids are nucleotides. Each nucleotide consists of three components: a phosphate group, a five-carbon sugar (deoxyribose in DNA and ribose in RNA), and a nitrogenous base. The four types of nitrogenous bases found in DNA are adenine (A), thymine (T), cytosine (C), and guanine (G). In RNA, uracil (U) replaces thymine.

DNA molecules have a double-stranded helical structure, with the two strands running in opposite directions and held together by hydrogen bonds between complementary base pairs. Adenine pairs with thymine (A-T), and cytosine pairs with guanine (C-G).

DNA Replication:

DNA replication is the process by which a cell duplicates its DNA to pass on genetic information to its daughter cells during cell division. This crucial process ensures that each new cell receives an identical copy of the genetic material.

The steps of DNA replication are as follows: 

  • Initiation: Proteins called DNA helicases unwind and separate the double-stranded DNA at specific sites called origins of replication. 
  •  Elongation: DNA polymerases synthesize new complementary strands along each separated template strand. The leading strand is synthesized continuously, while the lagging strand is synthesized in short fragments called Okazaki fragments. 
  • Termination: The replication process is completed when the entire DNA molecule is copied. The resulting two DNA molecules are identical to each other and to the original DNA molecule.

Protein Synthesis (Translation): 

Protein synthesis, also known as translation, is the process by which genetic information encoded in mRNA (messenger RNA) is used to build specific proteins. It occurs in ribosomes, where the genetic code is read and translated into a sequence of amino acids to form a functional protein.

The steps of protein synthesis are as follows

  • Initiation: The ribosome binds to the mRNA at the start codon (AUG), and the first tRNA (transfer RNA) carrying the amino acid methionine attaches to the ribosome. 
  • Elongation: The ribosome moves along the mRNA, reading each codon, and matching it with the appropriate tRNA carrying the corresponding amino acid. Peptide bonds form between adjacent amino acids, creating a growing polypeptide chain.
  • Termination: The ribosome reaches a stop codon (UAA, UAG, or UGA) on the mRNA, signaling the end of protein synthesis. The newly formed protein is released, and the ribosome dissociates from the mRNA.

Gene Transfer in Microorganisms:

Gene transfer in microorganisms refers to the process by which genetic material is exchanged between different microorganisms, promoting genetic diversity and adaptation to new environments. There are several mechanisms of gene transfer in microorganisms, including:

  • Transformation: Microorganisms can take up foreign DNA directly from their surroundings, integrating it into their genome. 
  • Conjugation: In this process, two microorganisms form a physical connection, and genetic material is transferred from the donor to the recipient cell through a structure called the pilus. 
  • Transduction: Bacteriophages (viruses that infect bacteria) can carry bacterial DNA from one host cell to another, leading to gene transfer. 
  • Transposition: Some genetic elements, such as transposons, can move within the genome of a single cell or between different cells, facilitating gene transfer.

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Assignment Activity 2 : Illustrate the mechanisms for regulation of gene expression in a prokaryotic system.

Prokaryotic organisms, such as bacteria, tightly regulate their gene expression to adapt to various environmental conditions and ensure efficient use of cellular resources. The primary mechanisms for gene expression regulation in prokaryotes are as follows:

Transcriptional Regulation: 

Transcriptional regulation controls the initiation of transcription, where RNA polymerase binds to the DNA and starts synthesizing RNA. Prokaryotes use specific DNA sequences called promoters and regulatory proteins called transcription factors to control gene expression.

  • Repressors: Repressor proteins bind to operator sequences near the promoter region, blocking RNA polymerase’s access to the gene and preventing transcription. 
  • Activators: Activator proteins enhance transcription by binding to specific sites and assisting RNA polymerase in binding to the promoter, thus increasing gene expression.

Post-transcriptional Regulation:

 Prokaryotic cells can control gene expression after transcription through various mechanisms that affect mRNA stability and translation efficiency.

  • Riboswitches: These are regulatory elements present in certain mRNA molecules that can directly bind small molecules (metabolites). Depending on the bound molecule, riboswitches can change their secondary structure, affecting mRNA stability or translation.
  • RNA-binding Proteins: Certain proteins can bind to specific mRNA sequences and influence their stability or translation.

Transational Regulation: 

Prokaryotes can control gene expression at the translation stage by modulating the efficiency of protein synthesis.

  • Ribosome Binding Sites: The accessibility of ribosome binding sites (Shine-Dalgarno sequences) on mRNA can be regulated, affecting translation initiation.
  •  Small Regulatory RNAs (sRNAs): These short RNA molecules can bind to mRNA and modulate translation by blocking or promoting ribosome access.

Post-translational Regulation: 

Proteins can be regulated after translation to control their activity, stability, or localization.

  • Protein Modification: Phosphorylation, acetylation, and other chemical modifications can activate or inactivate proteins. 
  • Proteolysis: Enzymatic degradation of proteins can regulate their half-life and activity.

Assignment Activity 3 : Display laboratory practical skills in following experimental procedures for microbial genetics.

The laboratory practical skills in microbial genetics involve performing experiments to study genetic elements, gene transfer, and gene regulation in microorganisms. Some typical procedures might include:

Transformation Experiment:

  • Preparation of competent bacterial cells. 
  • Introduction of plasmid DNA (foreign DNA of interest) into the competent cells. 
  •  Incubation and selection of transformed cells using antibiotic resistance markers. 
  • Analysis of transformation efficiency and verification of successful gene transfer.

Conjugation Assay: 

  • Set up mating experiments between donor and recipient bacterial strains. 
  • Selection of transconjugants on appropriate growth media containing selective markers.
  •  Identification and confirmation of successful gene transfer through phenotypic traits or molecular techniques.

Analysis of Gene Expression: 

  • Isolation of RNA from bacterial cells grown under different conditions. 
  • Reverse transcription of RNA into complementary DNA (cDNA). 
  • Quantitative real-time PCR (qPCR) to measure gene expression levels. 
  • Analysis of gene regulation based on the qPCR results.

Plasmid DNA Extraction:

  •  Extraction of plasmid DNA from bacterial cultures using commercial kits or standard protocols. 
  • Agarose gel electrophoresis to assess the quality and quantity of extracted plasmid DNA.

Protein Expression and Purification: 

  • Cloning of a gene of interest into an expression vector. 
  • Transformation of the recombinant plasmid into host cells for protein expression.
  • Protein purification using chromatography techniques. 
  • Analysis of purified protein using SDS-PAGE or other relevant assays.

Please note that these laboratory practical skills can be adapted based on the specific aims of the microbial genetics research or learning objectives. Always follow appropriate safety guidelines and standard operating procedures while conducting experiments in a laboratory setting.

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