Ohio University, Athens, Ohio, USA.
In this lecture, fundamental concepts in the area of molecular biology will be presented. These include biologys and biotechnologys central dogma; the human genome project; the discordance between human gene number and corresponding protein number; and gene cloning techniques. Also presented will be procedures used to determine gene number and location (Southern blotting) and levels of gene expression at the RNA (Northern blotting, reverse transcription/polymerase chain reaction, micro-array/gene chip) and protein (Western blotting, proteomics) level. Additionally, three examples describing the cloning of genes/cDNAs and production of the respective recombinant therapeutic proteins will be offered. Finally, functional genomic concepts and protocols will be discussed including production of transgenic and gene-disrupted (knocked out) animals as well as methods to down regulate gene expression using antisense, ribozyme, or small inhibitory RNAs. The lecture will stress the basics of the various protocols with clinical examples cited.