Question

You believe that the cytosolic protein p007 is involved in mediating growth hormone-stimulated proliferation in cells. You wish to establish a cell model using your cultured fibroblasts. Provide clear explanations to the following, each worth 2.5 pts each. a) Describe how you would transfect the growth hormone receptor DNA into your cells to create stable cell lines, and include a description of the benefits of using stable cell lines versus transiently transfected cell line. b) Describe how you would measure specific receptor binding of GH to your stable cell line. c) Describe how you would measure growth hormone-induced proliferation in your stable cell line. Make sure to include controls. d) Describe how you would use RNAi to demonstrate the requirement for p007 in growth-hormone stimulated proliferation. Make sure to include controls.

Answer 1

Answer:

Following are the answer to this question:

Explanation:

In option a:

There are many ways, which can be defined as follows:  

• It keeping cells "qualified," use heat shock and handle ions like Ca and ice incubating.
• It uses Micro-injection with DNA.  
• It uses the biolistics or even the process of gene-gun.  

The stable cell charts indicate simple and definitive outcomes when their relationship to specific results. Moreover, relative to temporary cell lines, findings are more reliable and error-free. These lines also easily break and much less.

In option b:

Radio tests including linking tests could be used  

The Ligand Binding Assays is an analysis, that is based on the binding of ligands to antibodies, or other macromolecules. Its existence and distance of formed ligand-receptor complexes are defined using a system of measurement that is usually electrical or fluorescent.

In option c:

The method for measuring the levels of cell growth is accessible and the  Another way is to measure the metabolism within the same cell. So many color pigments were also accessible that can permeate an organism as well as react with some proteins as well as other factors, and that can effectively identify a color final product.  

Throughout relation to the amount of metabolism exercise, cell proliferation could be evaluated by researching one or even more cell markings. The BrdU limited liability assessment is a very well-published illustration.  

Its specific antibody can detect BrdU and high BrdUincorporations levels are linked to high levels of cell proliferation.  Command cell without DNA receptor growth hormone.

In option d:

The cell growth will stop after each use of RNA interference ( RNAi). The p007 formation as well as the effect on growth caused by the intrusion will be a goal. If development is delayed effectively, the theory will be tested successfully to a cell without any of the required DNA. The RNAi is the free cell