20. You are designing a construct for delivery of your favorite gene's cDNA into mammalian cells...

 

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20. You are designing a construct for delivery of your favorite gene's cDNA into mammalian cells in culture. Your cDNA sequence is 300 base pairs long and you have chosen a circular plasmid (5 Kb - Kilo base pairs) to serve as the vehicle for transfection. The plasmid has two restriction sites in its MCS: EcoRI and HindIII. Your cDNA has sticky ends produced with EcoRI at each 5' and 3' end. a. Draw a schematic of the recombinant plasmid and label all the sequences (4 points) b. Which restriction enzyme (endonuclease) will you use to cut the plasmid for insertion of your cDNA into the plasmid and why? (1 point) c. How many nucleotides does your recombinant plasmid have now? (1 point) d. What technique will you use to deliver the recombinant plasmid into mammalian cells across the cell plasma membrane? What principle is the technique based on? (1 point) 20. You are designing a construct for delivery of your favorite gene's cDNA into mammalian cells in culture. Your cDNA sequence is 300 base pairs long and you have chosen a circular plasmid (5 Kb - Kilo base pairs) to serve as the vehicle for transfection. The plasmid has two restriction sites in its MCS: EcoRI and HindIII. Your cDNA has sticky ends produced with EcoRI at each 5' and 3' end. a. Draw a schematic of the recombinant plasmid and label all the sequences (4 points) b. Which restriction enzyme (endonuclease) will you use to cut the plasmid for insertion of your cDNA into the plasmid and why? (1 point) c. How many nucleotides does your recombinant plasmid have now? (1 point) d. What technique will you use to deliver the recombinant plasmid into mammalian cells across the cell plasma membrane? What principle is the technique based on? (1 point)

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a b HindIII will be used to cut the plasmid for insertion of the cDNA because the cDNA has sticky en... View the full answer