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Hi,
The jellyfish is an eukaryote and e.coli is a prokaryote. The jellyfish GFP gene has introns and Exxon’s , whereas the prokaryotes do not contain any introns. Also the jellyfish GFP promoter is eukaryotic which will not work in prokaryotic. Before cloning the gene in e.coli, the introns need to be removed and promoter needs to be changed. Best way is to use a mRNA of gGFP and reverse transcribe it and clone it using pBAD vector.
2.
The CTD of pol II is necessary for promoter recognition and binding. It has is necessary for successful initiation off transcription. Under the effect of toxins the RNA pol would find loosely to promoter. It will also result in abortive initiation.
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