26 Jun You are designing primers to clone the cystic fibrosis gene
You are designing primers to clone the cystic fibrosis gene, its promoter or both into an expression vector. You have a vector that already has a strong promoter driving the expression of the green florescent protein (GFP) gene below. You will modify this vector to create several different plasmids to suit the 4 scenarios described in the questions. In each case you need to decide whether you need to create a transcriptional fusion or a translational fusion. You should also consider whether you need to replace the promoter, the gene, or both. Below is the sequence of the cystic fibrosis promoter, the complete cDNA for the cystic fibrosis gene and the cDNA for the green florescent protein sequence. Within the GFP sequence pay particular attention to the location of the BamHI, Nsil, SreI and PvuI sites. To make this realistic, all primers that are designed must be 22 nucleotides long plus required restriction sites. Also to minimize the number of possible answers, use HindIII and EcoRI to exchange the promoter and the BamHI, NsiI, SreI and PvuI sites to make modifications to the gene.
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