By Michael J. Gibney, Ian A. Macdonald, Helen Roche
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Additional resources for Nutrition and Metabolism, 2nd Edition
Upon translation the different mRNA isoforms will give rise to different isoforms of the protein product of the gene. This is a relatively common phenomenon and although the physiological or metabolic relevance of the different isoforms of many proteins is not fully understood it may be relevant to molecular nutrition. For example, the peroxisome proliferator activator receptorgamma (PPARg) gene can produce three different isoforms of mRNA (PPARg1, PPARg2, PPARg3) as a result of different promoters and alternative splicing.
The intensities of the bands on the gels are analysed by densitometry, which is proportional to the concentration. Alternatively, it is also possible to use a labelled (radioactive, biotin or digoxigenin) probe, which is complementary to the amplified gene of interest. After gel electrophoresis, blotting and denaturation these probes hybridise with the PCR product, which can be visualised and quantified. Currently, most methods make use of intercalating agents (like cyber green) of fluorescent-labelled primers during the PCR in conjunction with PCR equipment with an optical detection system.
Thus, it is evident that a clear research question is needed before the choice of animal model can be made. Using molecular tools applied to the germ cells of animals, it is possible to insert an isolated (foreign) gene sequence into an animal’s genomic material. The resultant animals that carry such a foreign gene are called transgenic animals and have traits coded by these genes. Another approach is to knock out a certain endogenous gene. With this technique, a specific gene is modified, which results in a loss of function of that specific gene.