<P> In most cases, organisms containing recombinant DNA have apparently normal phenotypes . That is, their appearance, behavior and metabolism are usually unchanged, and the only way to demonstrate the presence of recombinant sequences is to examine the DNA itself, typically using a polymerase chain reaction (PCR) test . Significant exceptions exist, and are discussed below . </P> <P> If the rDNA sequences encode a gene that is expressed, then the presence of RNA and / or protein products of the recombinant gene can be detected, typically using RT - PCR or western hybridization methods . Gross phenotypic changes are not the norm, unless the recombinant gene has been chosen and modified so as to generate biological activity in the host organism . Additional phenotypes that are encountered include toxicity to the host organism induced by the recombinant gene product, especially if it is over-expressed or expressed within inappropriate cells or tissues . </P> <P> In some cases, recombinant DNA can have deleterious effects even if it is not expressed . One mechanism by which this happens is insertional inactivation, in which the rDNA becomes inserted into a host cell's gene . In some cases, researchers use this phenomenon to "knock out" genes to determine their biological function and importance . Another mechanism by which rDNA insertion into chromosomal DNA can affect gene expression is by inappropriate activation of previously unexpressed host cell genes . This can happen, for example, when a recombinant DNA fragment containing an active promoter becomes located next to a previously silent host cell gene, or when a host cell gene that functions to restrain gene expression undergoes insertional inactivation by recombinant DNA . </P> <P> Recombinant DNA is widely used in biotechnology, medicine and research . Today, recombinant proteins and other products that result from the use of DNA technology are found in essentially every western pharmacy, doctor's or veterinarian's office, medical testing laboratory, and biological research laboratory . In addition, organisms that have been manipulated using recombinant DNA technology, as well as products derived from those organisms, have found their way into many farms, supermarkets, home medicine cabinets, and even pet shops, such as those that sell GloFish and other genetically modified animals . </P>

What are the 3 steps for making a recombinant dna molecule