2018_19
Educational guide 
Faculty of Oenology
A A A 
english 
Bachelor's Degree in Biotechnology (2009)
 Subjects
  RECOMBINANT DNA TECHNOLOGY
   Contents
Topic Sub-topic
1. Introduction Introduction to genetic engineering to the basic concepts of cloning. Recombinant DNA; vectors and tools; incorporation of DNA to the host, basic techniques (PCR, electrophoresis ...).
2. Vectors for prokaryotes: plasmids and bacteriophages. Basic characteristics of plasmid and bacteriophage vectors. Selection. Improvement of the basic vectors. Plasmids pBR322 and pUC. Vectors derived from insertion and replacement lambda phage. Advanced vectors applications: Phage Display.
3. DNA manipulation. DNA purification. Cutting and binding: restriction enzymes, ligases, other enzymes used in genetic engineering.
4. Introduction of DNA in the host and selection of recombinants. Transformation by chemical and physical methods, calcium chloride method, electroporation. Virus. Identification of transformed organisms.
5. Genomic library Construction of genotypes.
6. Yeast cloning Cloning in S. cerevisiae. Vectors: chromosomal integration vectors (Yip) and autonomous replication vectors (Yep, YRp, YCp, YAC). Introduction of DNA in yeast. Expression of recombinant proteins. Yeast cell surface display. Applications: Yeast Two Hybrid, Yeast Three Hybrid.
7. Genetic engineering in animal cells. Transient and stable transfection. Transfection strategies. Selection methods. Vectors: non-replicating plasmids, plasmids with viral replicons, viral vectors (retroviruses, lentiviruses, adenovirus). Packaging cells. Protein expression. Gene silencing by interference RNA. CRISPR/CAS9 technique.
PRACTICAL SYLLABUS Cloning of the genome of the Lambda bacteriophage to the vector pUC18 of aplification to E. Coli.