Question

D 4. You want to use PGLO to produce red fluorescent protein (RFP) instead of green fluorescent protein (GFP). To do so, you cut out the GFP gene from PGLO with two restriction enzymes, and replace it with the coding sequence for RFP. Which two restriction enzymes will you choose to cut the RFP gene (out of a different plasmid) prior to ligating it into the PGLO plasmid? Select all that apply EcoRI EcoRV Ndel Psti Nhel D 3. You want to use PGLO to produce red fluorescent protein (RFP) instead of green fluorescent protein (GFP). To do so, you cut out the GFP gene from PGLO with two restriction enzymes, and replace it with the coding sequence for RFP. Which two restriction enzymes will you choose to cut the PGLO plasmid? Select all that apply EcoRI EcoRV Ndel Psti Nhel

Answer 1

Q.3. Based on the pGLO DNA sequence

EcoR I is on multiple cloning site located after GFP, so can be used.

EcoR V will make cut in araC so can not be used.

Nde I - two restriction sites are present on plasmid, so removing only GFP gene will not be possible, if we use this enzyme. Can not be used

Pst I is on multiple cloning site located after GFP, but another RE site is present on plasmid, which will affect bla gene. So can not be used,

Nhe I - plasmid pGLO has only 1 restriction site before GFP, so this enzyme can be used.

Answer: we can use EcoR I and Nhe I to cut the pGLO plasmid.

Q4. To cut RFP gene from other plasmid and ligate it to pGLO plasmid treated with EcoR I and Nhe I, we will need to use same restriction enzymes

Answer: use EcoR I and Nhe I to cut the other plasmid having RFP gene

Did this help? If yes, do give UP vote for this answer. Good day!