BMS415 Genetic Engineering & Bioinformatics (8)

• be able to describe the range of techniques in genetic manipulation, the cloning vectors available and the containment procedures required;
• be able to identify strategies for gene isolation, construction of libraries, DNA and peptide synthesis and DNA sequencing;
• abe able to demonstrate understanding of the complexities associated with engineering the expression of recombinant genes of medical or therapeutic importance, eg, human insulin, epidermal growth factor, human somatomammotrophin, human dystrophin genes;
• be able to describe the diagnostic, forensic, and medical applications of recombinant DNA technology.

The syllabus will cover the following topics: * review of basic molecular biology: theory, cloning vectors, mechanism of propagation; * tools of genetic manipulation: restriction endonucleases, methylases, nucleases, polymerases, ribonucleases, exonucleases, transcriptases, ligases, kinases. Uses of primers, adaptors, linkers; * cloning: cleavage of vector and insert, site compatibility, dephosphorylation of ends, ligation protocols and diagnostic techniques; * preparation of transformed cells: transformation and transfection techniques, routine screening procedures; * gene probing: hybridisation methods. Applications in biotechnology; * strategies for gene isolation: cloning DNA libraries and genomic DNA libraries, screening for genes of interest by hybridisation and immuno-detection methods. * Expression engineering: promoters, enhancers, homeo-boxes, fundamental regulatory elements. Secretion of engineered gene products; major factors affecting viability - refractile bodies, post-translational modification. Production, purification and analysis. Perspectives: development of expression vectors for mammalian cells; immunogenicity of engineered proteins - risk assessment in therapy; * DNA (Phosphite triester), protein engineering, oligonucleotides, peptide pharmaceuticals and automated synthesis. In vitro mutagenesis and elucidation of gene function *Genetic therapy: human germ-line and somatic gene therapy; transgenic livestock for industry; transgenic animals as pharmaceutical factories * DNA Sequencing: Sanger dideoxy, M13 and plasmid, troubleshooting, direct genomic sequencing with polymerase chain reaction. Perspectives: The Genome Sequencing Project; automated sequenators; * Scaling up mammalian and bacterial expression cultures: fermentation systems: batch, continuous and fed batch systems, turbidostat and chemostat cultures, downstream processing, inclusion bodies, secreted peptides, animal cell culture technology; * applications in diagnostics, forensics medicine and agriculture: peptide vaccines, peptide pharmaceuticals, vaccinia viruses, antibiotics, recombinant vaccines, food and beverage improvement.