Consensus prokaryotic promoter

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Your third task is to mutate the SID-1 promotor region. Below is 120 bp of the SID-1 genomic sequence ending with the start codon for the SID-1 ORF. A putative -10 equivalent in this promoter is the TATA sequence. You're interested to know if the -10 consensus sequence for prokaryotes could increase the expression of this eukaryotic gene.

A) What seven base pair primer pair could we use to mutate the current promoter to look more like a consensus prokaryotic promoter? Write the sequence and 5', 3' ends of your primers below.

B) You also want to PCR up this whole 120 bp sequence. You are told to order 10 bp PCR primers that are designed to bind to each end of this sequence. i) You object to using 10 bp sequences for the PCR experiment. Explain your reasoning.

ii) You're assured that the 10bp length will not be a problem. Given that guarantee, do you think this primer pair could work? Explain your reasoning.

Reference no: EM132412656

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