A strong constitutive promoter eg that from the ubiquitin...

A strong, constitutive promoter, e.g., that from the Ubiquitin gene, is needed in this case so that the herbicide resistant gene can be expressed in all parts of a plant. Assuming that only the coding region of the gene was known, which techniques would you use to clone the promoter? List two

Homework problem #6 6. In order to characterize the promoter region of your favorite gene (YFG), you have cloned a 200 nucleotide piece of genomic DNA upstream of the transcriptional start site and will use it to drive expression of the chloroamphenicol acetyltransferase (CAT) reporter in mammalian cells growing in culture. You have also deleted various portions of the 5' end of the cloned piece and have tested those constructs for CAT activity as well (left panel below). The results of CAT activity for constructs 1 through 5 are expressed in the graph (right panel below): For the following questions, answer A, B, C, D or any combination. a. What is the minimal promoter region (or regions) required for maximal expression? b. What region of the promoter contains a repressor binding site? c. What region of the promoter is dispensable for normal YFG regulation? d. In which region would mutations be most tolerated?

Sri K.

Question

A strong, constitutive promoter, e.g., that from the Ubiquitin gene, is needed in this case so that the herbicide resistant gene can be expressed in all parts of a plant. Assuming that only the coding region of the gene was known, which techniques would you use to clone the promoter? List two

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