Molecular Genetics

Code:

MI072111

Acronym:

GENMOL

Keywords
Classification	Keyword
OFICIAL	Natural Sciences

Instance: 2013/2014 - 1S

Active?	Yes
Responsible unit:	Biochemistry Laboratory
Course/CS Responsible:	MSc in Pharmaceutical Sciences

Cycles of Study/Courses

Acronym	No. of Students	Study Plan	Curricular Years	Credits UCN	Credits ECTS	Contact hours	Total Time
MICF	311	Official Curriculum	2	-	7	65	189

Teaching language

Portuguese

Objectives

Study of the basic mechanisms that allow the genetic DNA transmission based in replication, transcription, processing and translation processes. Modifications at DNA level analysis after recombination, mutation, transposition, transduction and viruses. Study of DNA repair mechanisms. Prokariotic and eukariotic gene expression regulation analysis to understand how gene expression is controlled and cells synthesize the right proteins at the right time in the right amounts. Study of the molecular mechanisms associated with cancer and related processes that involve the cell cycle regulation, mutations in proto-oncogenes and anti-oncogenes. The use of DNA replication technology as a dominant approach for studying the basic biological processes that allow cloning and identification of genes, analysis of genetic expression and regulation, diseases diagnosis, production of drugs and vaccines, and gene therapy.

Learning outcomes and competences

The advances that occur in Genetics, including the progress of scientific research, international engagement in mapping the human genome project (HGP) and development of specific techniques are crucial for the assessment of physiological and pathological systems in various areas of health but equally relevant for agriculture, chemical, electronics, energy, toxicology, pharmaceutical and forensic sciences. The knowledge acquired in this UC could be applied to several UCs which are then taught in the curriculum of the MICF, giving powers to the pharmacist for developing and implementing in various areas of work. The execution of laboratory work is done with the aim of carrying out a specific methodology, to contact directly with organic products, materials, reagents and instruments, as well as consolidate the improvement of scientific concepts.

 

Working method

Presencial

Pre-requirements (prior knowledge) and co-requirements (common knowledge)

Are required prior knowledge of Cell Biology, General Microbiology and Biochemistry and simultaneous knowledge of Organic Chemistry and Immunology.

Program

Tutorial program:
Desoxyribonucleic acid (DNA) - DNA structure and composition. DNA denaturation and renaturation. DNA packadge. Chromatina and chromosomes. DNA types.
Genetic code - Wobble pairing. The modified genetic code in mitochondria.
DNA replication - Different types of DNA replication
Transciption. Transcription regulation. Prokariotic and eukariotic RNA synthesis.
RNAs and RNA processing - Prokariotic and eukariotic messenger RNA, transfer RNA and ribosomal RNA
RNA editing. Pseudogenes.
Translation and protein synthesis - initiation, elongation and termination. Protein location.
DNA mutations - spontaneous and induced; frameshift; point mutations; by transposition; reversible; by suppression; ribosome mutations; suppression of nonsense mutation by tRNA mutants. Ames Test.
DNA repair - by fotorreactivation; by alkilation; by excision; by mismatch repair; by pos-replication or homologous repair; by SOS gene induction; by error prone; by GO system.
Genetic recombination - homologous; site specific and by transposition. DNA mapping.
Phage genetics - structure, infection, replication, recombination.
Bacteria - chromosome; mutation in E. coli; mutant enrichment; genetic recombination in E. coli: conjugation; transduction, transformation, transposition.
Recombinant DNA technology - cloning; restriction enzymes; restriction map; DNA polymerase I, DNA ligase, Reverse transcriptase; S1 nuclease; Terminal transferase; Exonuclease III; Cloning vectors: plasmids; l phage; PACs; BACs; YACs; expression vectors; Genomic DNA libraries, cDNAs libraries and expression libraries; Chromosome walking; DNA sequencing (Maxam-Gilbert, Sanger); DNA unidirectional deletions; Single stranded conformation polymorphism; Southern blot; Northern blot; Western blot; Chemical synthesis of oligonucleotides; Determination of the transcription initiation site (S1 nuclease and Primer extension); In situ hybridization; Fingerprinting; Mutagenesis in vitro; Radioisotopes; Transgenic and knockout animals; PCR and PCR applications.
Prokariotic gene expression regulation - Proteins that regulates the access of RNA polymerase to the promotor; Transcription regulation; Lactose operon, Arabinose operon; triptophane operon; Regulation of repairing and repressing systems; Regulation of transcription and translation termination systems; Phage l: life cycle.
Eukariotic gene expression regulation: signals of cellular control; levels of answers, and control mechanism (activating proteins, metylation, association to the nuclear matrix).
Vírus: Classification; Structure; Cell integration and exit; Replication strategies.
Viroides and Prions.
Oncogenes and anti-oncogenes - Molecular mechanisms of cancer induction. Retroviral oncogenes; Cellular oncogenes; Oncogene proteins: classification and characteristics; Anti-oncogenes. Apoptosis.
Cycle regulation - G1/S, G2/S and metaphase/anaphase (APC) transitions.
Laboratorial program:
1 – Bacterial kinetics growth. 2 – Bacteriophage infection.3 – Isolation of plasmidic DNA . 4 – Electrophoresis analysis of DNA digested with restriction enzymes. 5 – Restriction map construction from a recombinant plasmid. 6 – Discussion of practical work. Students reports evaluation.

Mandatory literature

Sambrook J, Fritsch EF & Maniatis T ; Molecular cloning - a laboratory manual, Cold Spring Harbor Lab. Press, USA, 1989
Sambrook J, Fritsch EF & Maniatis T ; Molecular cloning - a laboratory manual, Cold Spring Harbor Lab. Press, USA, 1989
Arraiano CM & Fialho, A M ; O Mundo do RNA, Lidel, 2007
Arraiano CM & Fialho, A M ; O Mundo do RNA, Lidel, 2007

Complementary Bibliography

Videira A ; Engenharia Genética: princípios e aplicações, Lidel, 2001
Videira A ; Engenharia Genética: princípios e aplicações, Lidel, 2001
Alberts Bruce 070; Molecular biology of the cell. ISBN: 0-8153-1620-8
Alberts Bruce 070; Molecular biology of the cell. ISBN: 0-8153-1620-8
Lodish Harvey 070; Molecular cell biology. ISBN: 0-7167-3706-X
Lodish Harvey 070; Molecular cell biology. ISBN: 0-7167-3706-X
LeWin Benjamin; Genes VII. ISBN: 0-19-879277-8
LeWin Benjamin; Genes VII. ISBN: 0-19-879277-8

Teaching methods and learning activities

 

a) Lectures - 3 hours / week; Laboratory classes - 2 hours / week, power-point presentation, the subjects taught in the theory are associated with the practical use of the knowledge imparted. b) Students are encouraged to make bibliographical research and consulting scientific articles related to the subjects taught. c) The laboratory work (2 hours/week) is made individually and according to the established schedule in which the techniques are applied that allow the identification and cloning of genes using the concepts and knowledge acquired in lectures d) The responsible teacher is available to meet students when requested.

 

 

.

Evaluation Type

Distributed evaluation with final exam

Assessment Components

designation	Weight (%)
Exame	80,00
Participação presencial	0,00
Trabalho laboratorial	20,00
Total:	100,00

Amount of time allocated to each course unit

designation	Time (hours)
Estudo autónomo
Frequência das aulas
Trabalho laboratorial
Total:	0,00

Eligibility for exams

Student attendance to the laboratory classes is mandatory. Those students whose attendance is lower than 3/4 of the foreseen classes are considered as without attendance.
- Attendance to the lectures is not compulsory.

Calculation formula of final grade

 

Two frequencies will be made, one in the week of mid-term evaluation and the other at the end of the semester. Students must have a minimum score of 8 values at each frequency (on a scale of 20 values) and the approval at this UC requires a final value of 9.5 (in the average of two frequencies), in addition to the classification obtained in the laboratory evaluation.
Students, who choose to make a mid-term evaluation by UC, will have initially the 1st test, and then have the laboratorial exam in the last week of classes, that requires a minimum of 9.5 values to go to the 2nd test or do the final examination or recourse.

The classification below 8 values in the 1st frequency prevents the student of going to the 2nd frequency, having however the right to do a recourse exam. Those that obtain f less than 8 values in the 2nd frequency remit the students for the recourse exam.
Students that not choose the mid-term evaluation will have to make the laboratorial exam, where they have to obtain a minimum of 9.5 values to make the final examination or recourse.

Laboratory evaluation includes: a) analysis of two scientific papers related with a laboratory technique in the field of Molecular Genetics (0.75 values) b) theoretical questions (0.75 values) c) bioinformatics (0,5 values); d) laboratory examination (2 values). Laboratory evaluation is the sum of the marks obtained in these four components, which are mandatory, and requires a minimum grade of 9.5 values for the student to apply to the second frequency or the final exam.

Students who have already had this frequency UC will, necessarily, to do the laboratory  exam in the last week of classes.

Nooral exams.

 

Examinations or Special Assignments

Students under a special regime, who are not obliged to attend laboratorial classes have to perform the final laboratorial exam. The approval in this component is mandatory to proceed with the final written evaluation exam.

Special assessment (TE, DA, ...)

- In accordance with the evaluation rules for FFUP.

Classification improvement

The student has the right to request the repeat of the final written exam tests to improve the mark, only one time, in the normal or resit period, following the period in which the approval in the discipline was obtained.
- The final subject classification will be the higher of those obtained in the two tests carried out.

Observations

Essential knowledge of Cellular Biology Lodish H, Berk A, Kaiser CA, Krieger M, Matthew P, Bretscher SA, Ploegh H & Matsudaira P (2007). “Molecular Cell Biology”. W. H. Freeman, 6th Ed.; Alberts B, Johnson A, Lewis J & Martin Raff M. (2008). “Molecular Biology of the Cell”. Garland Science, 5thEd.], as well as Biochemistry ([Nelson, D. L. & Cox, M. M. (2000) “ Leninger – Principles of Biochemistry” - Worth Publishers 3rd ed.]and Basic Microbiology [Sambrook, J., Fritsch E. F. & Maniatis T. (1989). “Molecular cloning - a laboratory manual”. 2sd Edition, Cold Spring Harbor Laboratory Press, U.S.A.].