Question

A biology 207 student sub cloned the promoterless, GFP gene from pCamR-GFP into pUC18-207 using a double digest. They used competent, prototrophic DH5aE. coli cells in their transformation protocol and plated the transformation reaction onto minimal medium that contained additional supplements to help them screen and select for their desired transformants, which are transformed E. coli cells that express the GFP gene. All experimental procedures were followed such as incubation times etc. Note that GFP encodes for a protein that fluoresces green. All you need to answer the questions that follow are the diagrams of the plasmids below and the information throughout this problem Pfl108 11715 Bsa l 1766 Sal 12177 AmpR Xmn I 2294 AlwN I 1217 ORI pUC18-207 2700 bp Ssp l 2501 Afl III 806 lacZ promoter Sap l 683 Nde l 183 MCS 454 396 EcoRI KpnBam HI Xbal Sal Pst I Hind IlIl Sac l Not I

Answer 1

if we cut this plasmid by NotI and SacI, it will give GFP plasmid.

The distance between NotI and SacI site is 6011-1995 = 4016 base pair.

So GFP is around 4000 base pair.

2-

3300 Remaing part of LacZ Sall of pUCi8 900 Sall of GFP LacZ Sall of MCS Ba 2500 Green - GFP gene

So correct answer is option F-Gel F.

3300 Sall of pUCi8 900 Sall of GFP Sall of MCS 2500