Please answer the following question Your PI would like you

Please answer the following question.

Your PI would like you to use electrophysiology [patch-clamp) to determine whether the transmembrane protein is EITHER a sodium ion channel OR a sodium/potassium pump. You have taken the cloned transmembrane protein mRNA and injected into Xenopus oocytes. Assume the oocytes are expressing the protein well on their surface. You design an experiment to test the function of the protein and you decide to use the whole cell recording technique. Answer questions a & b, below. a. What are the essential components of your solutions (besides water, what other components will you use---for example, ion(s), glucose, nucleotides, etc?) Explain your reasoning for each and explain how changing the components of your solutions will help you determine whether it is a channel or pump? b. Assume you are measuring current. How will the data differ if you are measuring the current for sodium channels versus sodium/potassium pumps? In other words, what do you expect your data to look like? Feel free to draw a mock graph(s) or data plot(s) if you like. If necessary you can scan in your drawing and upload via Canvas. One page maximum will be sufficient to answer question #2. Be concise and to the point!

Solution

Ques-1

The essential components of solutions required for the generation of an equivalent intracellular environment in vivo to examine xenopus oocytes using whole cell recording or patch clamping finally to unravel the nature of ion channels is whether a transmmebrane protein or not are \"isotonic sodium chloride, glucose, potassium chloride and calcium and magnesium & bicarbonate salts\". These salts should be in oxygenated environment to make oocytes alive for experimental purposes. The ratio of the components in the isotonic solution keep the cell membrane of xenopus oocytes alive and promote cell survive with no cell death or apotosis during the experiment or when applying sufficient voltage to cell. In this method an electrode using a large, tip which greater than 1uM with considerably sharp tip associated with very delicate surface used to spike cells associated ion channel proteins by pressing this electrode over the cell membrane. Here we need to apply suction in order to pull the plasma membrane inside the electrode tip finally leading to form a tight seal with the electrode. This system connected to the amplifier and voltage meter to get the spike s across the proteins.