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Category > Applied Sciences Posted 12 Aug 2017 My Price 5.00

cDNA from RNA

You have recently cloned a cDNA from RNA isolated from human brain tissue. The cDNA may encode a putative adrenergic receptor, based on its putative DNA and amino acid sequence homologies to known adrenergic receptors. You can assume it will be a new member of the beta adrenergic receptor family, called beta5. You are also given a drug X100 which has potent action (EC50=1 x 10-11nM) in vivo similar to beta adrenergic agonists, but which does NOT bind any of the known alpha or beta receptors. You may assume an active, radiolabelled form of X100 ([125I]-X100) is available.

The following questions #1-11 refer to what experiments you would perform to characterize and confirm that this cDNA clone codes for beta5, a new member of the adrenergic receptor family which binds all the same agonists and antagonists as the beta2 receptor subtype, and uniquely binds the drug X100. You stably express this cDNA in a cytomegalovirus-promoted expression plasmid in Chinese Hamster Ovary cells (CHO cells) after selection with antibiotic, but unfortunately, you can detect NOdetectable specific binding to [125I]-X100! You need to determine why you can not detect binding, and what you must do to successfully express this receptor in cultured cells and to determine its potential as a drug target for human therapies. Since you can not detect specific binding, what would be your reasons for doing each of the following?

 

1. transfect the expression plasmid into other cell types, including monkey COS cells and human embryonic kidney cells (HEK cells)? Explain your answer.

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SmartExpert
(118)
Status NEW Posted 12 Aug 2017 01:08 AM My Price 5.00

cDN-----------A -----------fro-----------m R-----------NA-----------

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file 1502529687-cDNA from RNA.docx preview (183 words )
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