Special molecular biotechnology
FCH-DA_SMBAk. rok: 2020/2021
The course focuses on the use of molecular biotechnology in the preparation of transgenic organisms for the isolation of biotechnologically important proteins. Attention is paid to the use of modern molecular biology and biophysical methods for their use in the production of significant proteins and the study of their interactions with DNA.
Doporučená nebo povinná literatura
Glick B. R., Pasternak J. J.: Molecular Biotechnology. ASM Press, Washington 2003. (EN)
J. Čechová, J. Coufal, E.B. Jagelská, M. Fojta, V. Brázda, P73, Like Its P53 Homolog, Shows Preference for Inverted Repeats Forming Cruciforms, PLoS One. 13 (2018). doi:10.1371/journal.pone.0195835. (EN)
Způsob a kritéria hodnocení
The results of the study are verified by colloquium (S.E.R. Art. 34 of the BUT Brno), ie a discussion about the subject matter connected with the preparation of a written work dealing with the specified topic of the subject in the range of about 10 text pages; the debate takes about 30 minutes.
Basic methods of recombinant DNA technology: isolation and characterization of DNA and RNA, restriction endonucleases and other enzymes, cloning, polymerase chain reaction, gene mapping, methods of transformation, transformation vectors - plasmids, phages, cosmids, YAC. Identification of recombinant DNA. Gene expression in microbial (bacteria, yeast) and animal expression systems, markers. Identification and purification of recombinant proteins (PAGE-SDS, Western blot, immunodetection).
• The relationship between chemical production and biotechnology
• Use of biophysical and molecular biological methods in DNA isolation and recombinant DNA technology
• Modern cloning methods, GATEWAY system
• Biotechnological use of genetically modified microorganisms
• Use of bioinformatic approaches in the study of nucleic acids and proteins
• Preparation of the transgene construct and protein isolation
The aim of the course is to introduce students how to use transgenic organisms in the preparation of useful products, especially proteins. Students will get ready a review on the topic and will proceed all steps necessary to isolate the transgenic protein using the GATEWAY cloning system.