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. |
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. |
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.
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