Question
Asked Oct 17, 2019
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A full-length eukaryotic gene is inserted into a bacterial chromosome. The gene contains a complete promoter sequence and a functional polyadenylation sequence, and it has wild-type nucleotides throughout the transcribed region. However, the gene fails to produce a functional protein.

a)List at least 3 possible reasons why this eukaryotic gene is not expressed in bacteria.

b)What changes would you recommend to permit expression of this eukaryotic gene in a bacterial cell?

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Expert Answer

Step 1

The eukaryotic DNA is composed of coding and non-coding sequences, almost 90% are non-coding (introns) region for proteins. The bacterial genome consists of only exons (coding sequences). Also, the expression in bacteria and in eukaryotes varies in many aspects.

Step 2

a) To initiate the transcription in bacteria, there is a need of the specific promoter region called a Pribnow box and -35 consensus sequences, whereas eukaryotes need TATA box and many transcriptional factors in order to initiate the transcription. Because of these specific promoter regions, RNA polymerase cannot bind to the eukaryotic promoter and cannot start transcription and translation.

Absence of Shine-Dalgarno sequence in eukaryotes does not transcribe the gene inserted into bacteria.

The gene possibly ...

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