For questions 5 through 9: You have isolated an epithelial cell line which is unresponsive to...

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Biology

For questions 5 through 9: You have isolated an epithelial cellline which is unresponsive to EGF (normal epithelial derived celllines require EGF in the culture media for growth). You suspectthat EGF receptor signaling in this cell line is impaired, and youdesignate this a mutant cell line. As an assay for EGFR signaling,you have stably transfected this mutant cell line and a normal cellline with a luciferase-based reporter construct that is responsiveto the transcription factor Elk-1. You want to determine whatcomponent of the signaling pathway is impaired. 5. (5 pts) Brieflydescribe a method you could use to transfect the luciferasebasedreporter construct DNA into a cell, and then describe how you couldestablish (or select for) a stable cell line expressing thisreporter construct.

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In order to create stable cell line which expresses luciferase construct we need to clone the luciferase gene in the pCDNA31 plasmid This plasmid has neomycin antibiotic resistance gene Before See Answer

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