From Single Gene Studies to Functional Genomics I. Recombinant DNA technology allows any DNA sequences to be tested A. DNA cloning techniques allow DNA molecules of any sequence to be assembled. Typically, recombinant DNA molecules are assembled in plasmid vectors and propagated in E. coli since this system is one of the simplest for routine manipulations and analysis of products. Molecules assembled in E. coli plasmid vectors can then be transferred to specialized vectors that may be less convenient to use than E. coli plasmid vectors. Automated DNA synthesizers can readily synthesize sequences up to 100 to 150 nucleotides and these sequences can be incorporated into recombinant molecules as well.
Most manipulations make use of restriction enzymes.
PCR can be used to modify the ends of amplified fragments to facilitate assembly of recombinant molecules. B. Often, the goal of cloning a gene or cDNA is to study the protein it encodes. In vitro transcription and translation can be used to produce proteins of interest. Expression vectors allow the production of very large amounts of proteins encoded by the sequences they contain. 1. reporter genes code for proteins that can easily be detected in the system being studied.