Genetic Engineering of Microorganisms
Code: 101977Credits: 6
| Degree programme | Type | Course |
|---|---|---|
| Genetics | OP | 4 |
Contact lecturer
- Name :
- Jesús Aranda Rodriguez
- Email :
- jesus.aranda@uab.cat
Teaching staff
- Sergi Torres Puig
- Jordi Corral Sabado
- María Perez Varela
- Susana Campoy Sanchez
Group languages
You can consult this information at the end of the document.
Prerequisites
It is recommendable to have studied or are studying Molecular Biology of Prokaryotes.
Objectives
The main objective of this course is that the student will be able to design procedures for the genetic manipulation of microorganisms.
Therefore during the development of the subject, the student must reach the following capacities:
• To know how to identify different types of microbial vectors, recognize their applications and design new ones
• To know how to apply methodologies and strategies of cloning
• To recognize the implication of the characteristics of each microorganism (immunity systems, recombination capacity, codon usage, etc.) in the proposed experimental design
• to know how to choose the most appropriate genetic transfer technique in each proposed case
• To be able to design efficient strategies for obtaining, enriching and selecting mutants
• To know how to build gene fusions and recognize their possible applications
• To recognize the main characteristics of potential bacterial targets for drugs, vaccines, and diagnostic reagents development.
Learning outcomes
- CM24 (Evaluate the validity and significance of experimental results on molecular mechanisms and developmental processes.) Evaluate the validity and significance of experimental results on molecular mechanisms and developmental processes.
- CM25 (Design basic research projects aimed at the study of gene regulation and mobile genetic elements in prokaryotes and eukaryotes.) Design basic research projects aimed at the study of gene regulation and mobile genetic elements in prokaryotes and eukaryotes.
- CM26 (Adapt the communication of complex concepts of molecular biology, pathogenesis and development to the needs of different audiences.) Adapt the communication of complex concepts of molecular biology, pathogenesis and development to the needs of different audiences.
- KM18 (Describe the mechanisms of gene expression and its regulation in prokaryotes and eukaryotes, integrating their bioethical implications and the gender perspective in genomics.) Describe the mechanisms of gene expression and its regulation in prokaryotes and eukaryotes, integrating their bioethical implications and the gender perspective in genomics.
- KM19 (Explain the molecular processes of the cell cycle, apoptosis, and the genetic basis of cell development and differentiation.) Explain the molecular processes of the cell cycle, apoptosis, and the genetic basis of cell development and differentiation.
- SM22 (Interpret the genetic basis of phenotypes and diseases based on the mechanisms of gene regulation and development, considering the impact of differences by sex.) Interpret the genetic basis of phenotypes and diseases based on the mechanisms of gene regulation and development, considering the impact of differences by sex.
- SM23 (Relate the structure and dynamics of nucleic acids to the application of molecular techniques in the study of developmental biology.) Relate the structure and dynamics of nucleic acids to the application of molecular techniques in the study of developmental biology.
- SM24 (Apply experimental design strategies and hypothesis formulation for the study of gene regulation processes and molecular biology.) Apply experimental design strategies and hypothesis formulation for the study of gene regulation processes and molecular biology.
Contents
The content of the course consists of the following topics:
Unit 1. DNA introduction systems in bacteria. Natural transformation in gramnegative and grampositive bacteria. State of competence. Molecular mechanisms associated with natural transformation. Induced transformation. Electrotransformation. Design and optimization of transformation systems in bacteria lacking natural transformation. Other Systems of DNA transference.
Unit 2. DNA vectors and cloning strategies in bacteria. Requirements of cloning vectors. Expression vectors. T-type vectors. Mobilizable vectors. Suicide vectors. Shuttle vectors. Integrational vectors. Genetic characteristics of vector accepting cells. Construction of DNA libraries in vitro and in vivo. Cloning by complementation: anabolic or catabolic genes. Regulatory gene isolation methods. Obtaining virulence genes. Cloning of toxic genes.
Unit 3. Bacterial gene fusions. Transcriptional and translational fusions. Gene fusions in polycistronic units. Fusion vectors: general characteristics. Random gene fusions. Methods for the construction of gene fusions. Construction of gene fusions by PCR, OE-PCR and Gibson assembly. Applications and examples of gene fusions.
Unit 4. Mutagenesis in bacteria. Random mutagenesis in vivo. Use of chemical or physical methods. Criteria and methods for the selection and enrichment of mutants. Transposons. Minitransposons. Plasposons. Transposomes. Methods for the identification and confirmation of mutants. In vitro mutagenesis of cloned genes.
Unit 5. Gene substitution in bacteria and generation of knockouts. Obtaining mutants by gene disruption and by gene substitution. Lambda Red system. Obtaining scarless mutants. Counter selection systems. I-SceI system. Use of CRISPR/Cas9 technology to obtain mutants. Methods for the identification and confirmation of mutants. Systems for the reintroductionof altered genes in the bacterium of origin. Insertion into the chromosome of new genes or constructs.
Learning activities and methodology
| Title | Hours | ECTS | Learning outcomes |
|---|---|---|---|
| Study and other autolearning activities | 50 | 2 | CM24, SM22, SM23 |
| Tutorship | 1 | 0.04 | KM18, KM19 |
| Preparation of posters and questionnaires | 34 | 1.36 | CM24, CM25, CM26, KM18, KM19, SM22, SM23, SM24 |
| Participatory master classes | 30 | 1.2 | CM24, CM25, CM26, KM18, KM19, SM22, SM23, SM24 |
| Reading recommended texts | 20 | 0.8 | |
| Seminars | 12 | 0.48 | CM24, CM25, CM26, KM18, KM19, SM22, SM23, SM24 |
The course is organized in two modules:
Theoretical module: where participatory master classes are combined with problem-based learning sessions where theoretical concepts are worked through the resolution of practical cases.
Seminar module: in which through collaborative learning, students work on different aspects of actual experimental designs present in recent scientific articles. At the beginning of the course, students choose, following the guidelines set by the teaching staff, a scientific article related to the field of genetic engineering of microorganisms from which they make a poster. The schedule of activities like classroom work sessions, exhibition, and discussions, as well as the delivery dates of the proposed activities will be defined at the beginning of the course by the teachers.
Assessment
Continuous assessment activities
| Title | Weight | Hours | ECTS | Learning outcomes |
|---|---|---|---|---|
| team-work or individual self-evaluation | 5% | 0 | 0 | CM24 |
| Discussion and participation in the classroom | 10% | 0 | 0 | KM19, SM22 |
| Classroom and virtual classroom submissions | 10% | 0 | 0 | CM24, CM25, CM26, KM18, KM19, SM23, SM24 |
| Poster | 25% | 0 | 0 | CM24, CM25, CM26, KM18, KM19, SM22, SM23, SM24 |
| Written test (resolution of practical cases) | 40% | 3 | 0.12 | CM24, CM25, CM26, KM18, KM19, SM22, SM23, SM24 |
| Resolution of questionnaires | 10% | 0 | 0 | CM24, CM25, CM26, KM18, KM19, SM22, SM23, SM24 |
Seminar module evaluation
The evaluation of the seminars is done through the evaluation of different activities related to a scientific article
Theoretical module evaluation
This activity is evaluated through a written individual exam that accounts for 80% of this module.
During the theory module sessions, evaluation activities may be carried out, contributing up to 20% of this module.
To pass this module, it is necessary to obtain a score of 5 points or higher in the written individual exam.
If the grade obtained is less than 5, the student must take the retake examination.
To be eligible for the retake process, the student should have been previously evaluated in a set of activities equaling at least two-thirds of the final score of the course or module.
Students who have passed the module may submit to a grade improvement test waiving the grade obtained previously in the individual written exam. The scheduled date for the second chance test is that of the second chance examination. Students wishing to take the grade improvement test must communicate it by mail to the teacher responsible for the subject at least 72 hours before the day scheduled for the second chance examination.
The final grade of the course will be theaverage of the grades obtained in both modules, being necessary to have passed separately each of them.
The student will be graded as \"Non-evaluable\" if the weight in of all conducted evaluation activities is less than 67% of the final score.
Single assessment
The evaluation of the theoretical module consists of a single test that will be the same as that of the type of continuous assessment, this test will account for 50% of the final grade for the subject and the same system of evaluation will be applied for recovery than for continued evaluation.
The evaluation of the activities of the seminar module will mean 50% of the final grade for the subject. The students who take advantage of the single evaluation may deliver all the evidence together (including the oral presentation) on the same day as the one set for the synthesis test. The single assessment test will be carried out coinciding with the same date set in the calendar for the last continuous assessment test.
Uses of AI
For this subject, the use of Artificial Intelligence (AI) technologies is permitted exclusively in support tasks, such as bibliographic or information fencing, text correction or translations, etc. It takes time to clearly identify which parts have been generated by this technology, specify the aspects and include a critical reflection on how these have influenced the process and the final result of the activity. The non-transparency of the use of AI in this evaluable activity is considered a lack of academic honesty and may entail a partial or total penalty in the activity grade, or major sanctions in serious cases.
The commission of any irregularity in an evaluation activity (academic fraud, plagiarism, or improper use of AI, unless such use is expressly authorized in the course guide), which may lead to a significant variation in the grade, means that this activity will be graded with a 0. In the event that the course guide stipulates that obtaining a minimum grade in this evaluation activity is an essential requirement to pass the course, or if several irregularities occur in the evaluation activities of the same course, the final grade for the course is 0. Regardless of this, a disciplinary proceeding may be initiated against the student who incurs any of these irregularities.
Bibliography
As reference bibliography of basic concepts it is recommended:
Larry Snyder i Wendy Champness. Molecular Genetics of Bacteria (3rd or 4th Edition). ASM press (ISBN: 978-1-55581-399-4 and ISBN:978-1-55581-627-8).
https://bibcercador.uab.cat/permalink/34CSUC_UAB/15r2rl8/cdi_askewsholts_vlebooks_9781118685112
Jeremy W. Dale and Simon F. Park. Molecular Genetics of Bacteria, (5th Edition) Wiley-Blackwell (ISBN: 978-0-470-74184-9).
Other recommended texts as well as links of interest will be available in the Moodle classroom of the subject.
Software
ot applicable.
Course groups and languages
The information provided is provisional until November 30. After this date, you will be able to consult the language of each group through this link. To access the information, you will need to enter the course CODE