A biology 207 student sub cloned the promoterless, GFP gene from pCam*-GFP1 207

Question

A biology 207 student sub cloned the promoterless, GFP gene from pCam*-GFP1 207 using a double digest. They used competent, prototrophic DH5aE. coli cells Time left 2 transformation protocol and plated the transformation reaction onto minimal m 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. Bsal 1766 P1l108 1 1715 Sal 12177 AmpR Xmn I 2294 AlwN I 1217 ORI pUC18-207 -2700 bp Ssp / 2501 Afl III 806 lacZ promoter Sap / 683 Nde l 183

Explanation / Answer

1. The student used Hind III and SacI as these two RE sites are not present wihin the cloning material hence there is no chance of it being digested.

2. IPTG and Histidine are used for selection of transformed colonies.

3. Flouresces green and white coloured colonies.

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