Microsoft PowerPoint - INTRODUCTION TO Recombinant DNA

Microsoft PowerPoint - INTRODUCTION TO Recombinant DNA - r...

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Unformatted text preview: r INTRODUCTION TO INTRODUCTION TO INTRODUCTION TO INTRODUCTION TO Recombinant DNA Recombinant DNA Recombinant DNA Recombinant DNA By By By By S. M. S. M. S. M. S. M. Rafigh Rafigh Rafigh Rafigh 2011 2011 2011 2011 I. Introduction A. The Central Dogma • Gene Expression – DNA makes RNA makes protein • DNA is transcribed into mRNA (nucleus) which is translated into protein (cytoplasm) – Transcription of genes relies on the interaction of DNA binding proteins with the regulatory elements of the gene (promoter) • Some genes are activated in all cell types ousekeeping genes) and are regulated by (housekeeping genes) and are regulated by ubiquitous promoters) • Some genes are activated in a restricted manner (tissue-specific genes) and are regulated by tissue- specific promoter and enhancer sequences – Pre-mRNA matures by splicing (the removal of intervening sequences) and polyadenylation (the addition of poly A tracts to the 3’ end), processes believed to stabilize mRNA and assist in its transport to the cytoplasm – Some mRNA transcripts are differentially spliced giving rise to proteins which may have different functions – Translation of mature mRNA occurs in the cytoplasm and involves the interaction with tRNA and rRNA complexes • Codons dictate where translation starts (AUG), stops AA, UAG, UGA), and which amino acids to (UAA, UAG, UGA), and which amino acids to incorporate into the growing polypeptide chain • Any mutations which have resulted in the addition or removal of nucleotides can alter the reading frame resulting in the translation of non-sense proteins or premature termination of translation. • Advances in Molecular Biology – The combination of restriction/modification enzymes and hybridization techniques enable the application of a wide variety of procedures B. Applications • Gene isolation/purification/synthesis • Sequencing/Genomics/Proteomics • Polymerase chain reaction (PCR) • Mutagenesis (reverse genetics) xpression analyses (transcriptional and translational levels) • Expression analyses (transcriptional and translational levels) • Restriction fragment length polymorphisms (RFLPs) • Biochemistry/ Molecular modeling • High throughput screening • Combinatorial chemistry • Gene therapy • Recombinant Vaccines • Genetically modified crops • Biosensors • Monoclonal antibodies ell/tissue culture • Cell/tissue culture • Xenotransplantation • Bioremediation • Production of next generation antibiotics • Forensics • Bioterrorism detection C. Definition of recombinant DNA • Production of a unique DNA molecule by joining together two or more DNA fragments not normally associated with each other NA fragments are usually derived from • DNA fragments are usually derived from different biological sources D. Development of molecular biology • Early research on prokaryotic genetics and the development of molecular techniques has led to a new discipline called MOLECULAR IOLOGY BIOLOGY...
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This note was uploaded on 06/21/2011 for the course CHE 774 taught by Professor Prof.rafigh during the Spring '11 term at Sharif University of Technology.

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Microsoft PowerPoint - INTRODUCTION TO Recombinant DNA - r...

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