MBLG2071/2971

Molecular Biology and Genetics A

Course Information

These course outlines are a guide only. They are provided for the information of prospective students. Although every effort is made to ensure the most up to date information is provided, timetables often change each semester due to the availability of rooms and resources. Content (including lecture/practical topics, assessment and textbooks) is also regularly reviewed to ensure relevance and effective learning.

Unit of Study Overview

MBLG2071 extends the basic concepts introduced in MBLG1001. It provides a firm foundation for students wishing to continue in the molecular biosciences as well as for those students who intend to apply molecular techniques to other biological or medical questions. MBLG2971 is the advanced option of the course.

The theory component is presented in 23 lectures (2 per week). It explores the regulation of the flow of genetic information in both eukaryotes and prokaryotes. The central focus is on the control of replication, transcription and translation and how these processes can be studied and manipulated in the laboratory. Experiments in model organisms are provided to illustrate how the field has advanced, together with discussion of work carried out in human systems and the relevance to human genetic diseases. The tools of molecular biology are taught within the context of recombinant DNA.

The practical course (6 four hour sessions; one every fortnight) focuses on the fundamental techniques in molecular biology research, building on the skills learnt in MBLG1001. Specifically students will: use spectrophotometry to identify and quantify nucleic acids and colourless biochemicals, explore the lac operon system to investigate regulation of gene expression, perform PCR analysis on their own DNA and isolate and analyse plasmid DNA. As with MBLG1001, strong emphasis is placed on the acquisition of generic and technical skills.

The advanced course (MBLG2971) covers the same lecture material as the normal (MBLG2071) course. The practical course, however, contains a number of different laboratory experiments designed to cover some of the material in more depth.

Course Coordinator Contact Details

Dr Markus Hofer
Room: 705
Phone: 9351 2233
E-mail:

Prerequisites

For MBLG2071: MBLG1001 and 12cp of Junior Chemistry

For MBLG2971: Distinction in MBLG1001 and 12cp of Junior Chemistry

Timetable

1st Lecture: Tuesday 11:00am Merewether Lecture Theatre 2 (Rm 136)
2nd Lecture: Friday 11:00am Carslaw Lecture Theatre 157

Practicals: Each student attends 6 practical sessions, one per fortnight. Sessions are held from 1-5pm, Tue, Thur or Fri in Room 380, Molecular Bioscience Building

Textbooks

Watson et al Molecular Biology of the Gene, 7th Edition.
Also suitable are any of the recent Biochemistry Textbooks that are all entitled ‘Biochemistry’ but identified by their different authors: e.g. Garrett & Grisham, Voet & Voet or Stryer. Any of these textbooks would be a good alternative for those wishing to continue with Biochemistry in second semester.

Lecture Outlines

Please note that this list is meant as a GUIDE ONLY. The order and content of each lecture may vary from that described and this list may not adequately reflect the emphasis put on each topic by each lecturer.

  1. Transcriptional Regulation I: Models of Gene Expression: the Lac Operon. Uses of the beta-galactosidase in Molecular Biology, blue/white colour selection.
  2. Transcriptional Regulation II: Models of Gene Expression: the Trp Operon. Transcriptional and translational control (attenuation).
  3. Eukaryotic transcriptional Regulation: Promoters, RNA polymerases and transcription factors. Basal transcription, elongation, enhancers.
  4. Post-transcriptional Processing of Splicing: (alternative splice sites), cleavage, poly-adenylation, transport, stability of mRNA, control of aberrant transcripts RNA interference.
  5. Translation in Eukaryotes: Extending your knowledge of prokaryotic translation from MBLG1001, we learn about the similarities and differences regarding translation in eukaryotes.
  6. Translational Regulation I: Pre- and post-translational control. Control of translation in eukaryotes: alternative start sites, Iron-responsive elements, Control of protein stability and degradation.
  7. Translational Regulation II: Protein Trafficking: Transport of protein from the cytoplasm to mitochondria and nucleus. The secretory pathway through the endoplasmic reticulum and Golgi to the cell surface.
  8. Introduction to the structure of the Genome: How is DNA packaged in a cell? Chromosome length and diversity, differences between eukaryotic and prokaryotic chromosomes, packaging proteins e.g. histones and the packaging to a chromosome. Heterochromatin and euchromatin and their relationship to transcriptional regulation.
  9. Genomic sequence complexity: Size and complexity, C-value paradox, melting and annealing DNA, CoT curves and different classes of DNA. Introduction to the complexity of the genome: protein-coding portions of the human genome and non-coding RNA.Transcription of rRNA and tRNA by RNA pol I and III. Introduction to other non-coding RNAs.
  10. Eukaryotic replication: Starting with the E. coli replication fork covered in MBLG1001, initiation and termination in bacteria. Eukaryotic replication with multiple, linear chromosomes, the eukaryotic cell cycle, the link between DNA replication and the cell cycle; both prokaryotic and eukaryotic. Mitotic cell division revisited (prophase, metaphase etc).
  11. Cell cycle regulation: Cell cycle in eukaryotes, check points in the cell cycle, introduction to immortal cell lines, the constraints normal cells face with cell division, Meiosis review.
  12. Homologous recombination and meiosis: Leading on from the concept of cross-over covered by Dr Lyons in MBLG1001....What is happening at a molecular level? Models of homolgous recombination, recombination in eukaryotes, genetic consequences.
  13. Model Eukaryotes: Yeast, nematodes, fruit flies and mice in the service of molecular biology and genetics.
  14. Transgenesis: Methods of producing transgenic animals, conditional versus non-conditional transgenics.
  15. Gene Therapy: Applications of Gene Therapy, somatic vs germline, methods for introducing DNA into cells, viral mediated gene transfer, human gene therapy.
  16. Knockout Animals: Basic knockouts, Specialized knockouts, genome wide knockouts, Disease modelling.
  17. Techniques to measure gene expression: There is a difference between genome (DNA, which is the same in nearly every cell) and expressed genes (RNA and protein), which give rise to different cell types. How do we measure gene expression? Techniques for detecting and quantifying mRNA : (eg RNA isolation, Northerns, microarrays, Real time PCR, RNase protection assays), and protein (eg Western Blots, ELISAs, enzyme assays).
  18. Working with DNA experimentally: How different DNA polymerases (eg Klenow and Taq) are used in molecular biology: producing labelled oligonucleotide probes for Northern and Southern blots, for PCR (polymerase chain reaction) and DNA sequencing.
  19. DNA Repair: Importance of repairing the genome, repair of DNA damage, direct reversal (photoreactivation, methyltransferase), how deamination of cytosine is corrected.
  20. Mutation I: The need for change. Sources of error: replication errors (nature of mutations, tautomers of bases, escape of proofreading), mutation rates, spontaneous DNA damage (hydrolysis and deamination).
  21. Mutation II (Continued): environmental DNA damage (alkylation, oxidation and radiation), mutagenic DNA damage (base analogues and intercalating agents). Types of errors (insertion, deletion, substitution).
  22. Stem Cells: Totipotency, pluripotency,multipotency, embryonic and adult stem cells, nuclear transfer and cloning.
  23. Signalling pathways that control gene expression: Overview of the major receptor classes-cell surface and cytoplasmic, receptor coupled-signal transduction pathways, down-modulation and negative regulation of receptor signalling.

Practical Course

  1. UV Spectrophotometry: Introduces students to the use of UV spectrophotometry to identify and quantify bases and nucleic acids and biochemicals.
  2. Gene Expression I: Investigates regulation of the lac operon in E. coli, using IPTG, lactose, glucose and protein synthesis inhibitors.
  3. Gene Expression II: Students design their own experiment to examine a particular aspect of the lac operon system.
  4. DNA Fingerprinting I: Specific loci in students’ DNA amplified by the polymerase chain reaction.
  5. DNA Fingerprinting II & Plasmid Isolation: Gel electrophoresis analysis of PCR products and miniprep isolation of plasmid DNA.
  6. Plasmid Analysis: Gel electrophoresis analysis of plasmid isolates.

Assessment

Lecture course: 50% (end-of-semester examination).

Practical course: 50% (25% in-semester practical work, 25% end-of-semester examination).

Tasks: One 2.5 hour exam including ‘Theory’ and ‘Theory of Practical’ questions, 3 in-semester hand-in laboratory assignments, continuous assessment during practical classes and laboratory book hand-in for final assessment.