In biotechnology, researchers are engineering Escherichia coli to produce a plastic-degrading enzyme, PETase, originally identified in the bacterium Ideonella sakaiensis, to assist in the bioremediation of polyethylene terephthalate (PET) waste.
Name the class of enzyme used by bioengineers to cleave the PETase gene from donor DNA and to open the plasmid vector, and explain why using the same enzyme for both is essential.
Identify the enzyme used to join the sugar-phosphate backbones of the PETase gene and the open plasmid vector, and state the type of bond that it forms.
Instead of isolating the PETase gene directly from genomic DNA (which contains introns that bacterial host cells cannot splice), scientists often extract mature mRNA. Describe how this mRNA is converted into double-stranded DNA so that it can be incorporated into the bacterial plasmid vector.
Explain how an antibiotic resistance marker gene, such as the ampicillin resistance gene (blablabla), is used to identify host bacteria that have successfully taken up the recombinant plasmid.
