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

Question

A biology 207 student sub cloned the promoterless, GFP gene from pCam -GFP into pUC18-207 using a double digest. They used competent, prototrophic DH5??. 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 problenm Pfl108 1 1715 Bsa 1766 Sal 12177 Xmn I 2294 AlwNI 1217 ORI pUC18-207 2700 bp Ssp l 2501 Afl III 806 lacZ promoter Sap 683 Nde l 18:3 MCS 454 396 EcoRI KpnBam HI Xbal Sal 1 Pst I Hind Ill Sac Notl PstI 7000 EcoR 500 CamR ORI Bam HI 1500 Hind III 6034 Sal 1963 Bam HI 6029 Not I 6017 pCamR-GFP 8000 bp Xba l 1972 Bsa 1982 ?Notl 1995 Sac I6011 GFP

Explanation / Answer

I believe the correct answers to be

Answer 1) A) Xba1 and I) Hind III

Answer 2) A) IPTG, B) Ampicillin and C) X gal.

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