Question

1. The serum amyloid A (SAA) superfamily comprises a number of genes and proteins characterized from a range of mammalian species. These proteins are dramatically induced during the acute-phase injury response, suggesting an important short-term beneficial role in the response to tissue injury and inflammation. However, important disease associations have also been proposed for certain SAAs during chronic inflammation. Serum amyloid A contributes to a variety of conditions wherein normally soluble proteins become insoluble and are deposited in the extracellular space of various organs or tissues, disrupting normal function. The disease state, amyloidosis, occurs in various mammalian species including humans, as well as domestic and wild animals of Nature's list of vulnerable The cheetah (Acinonyx jubatus) is an endangered animal included on the International Union for Conservation species. The principal problems in propagation of the cheetah are inbreeding depression and low genetic diversity (O'Brien et al. 1985). Further exacerbating the problem, amyloid A (AA) amyloidosis is increasingly becoming an important cause of morbidity and mortality in captive cheetahs (Papendick et al. 1997). na is a big cat specialist and consults for several zoos. This year she is interested in the learning more about amyloidosis in cheetahs. By comparing the sequences of the SAA genes among human, mouse, hamster, rabbit, dog, cow, sheep, and horse, she designed primers that different species such as cheetah. Outine how Fontana should set up her PCR to amplify the cheetah SAA gene. us to conserved regions of the genes. Such primers are likely to be successful for the amplification of the corresponding gene in a

Answer 1

For PCR, there are three steps that need to be done.

1) Denaturation: In this step, the bonds of the sample DNA are broken to form single stranded genetic material. This takes place by heating it to 94 degree Celsius.

2) Annealing : The temperature is suddenly lowered to 52 degree Celsius to enable the primers to attach to the template DNA strand.

3) Extension : In this, the temperature is raised and the new strand of DNA is produced by an enzyme called Taq polymerase . The temperature required for this step is 74 degree Celsius. The enzyme Taq polymerase is obtained from a bacteria called thermus aquaticus.

These steps are repeated a number of times so that the required gene gets amplified.

Hope this helps :)