Recombinant DNA Technology: Creating Plasmid Vectors

The tetR gene acts as a reporter gene and has a specific recognition site to one of the restriction endonucleases used in step 2.

EcoRI and BamHI

The insulin subunit genes were produced in a laboratory, and the extra codon coding for methionine was added to facilitate the gene expression.

<p>To allow some of the recombinant plasmids to be taken up by the bacteria.</p> Signup and view all the answers

<p>To determine which bacteria successfully took up plasmids.</p> Signup and view all the answers

<p>DNA ligase is used to reestablish the sugar-phosphate backbone and create two different recombinant plasmids.</p> Signup and view all the answers

<p>To determine which colonies contain bacteria that took up recombinant plasmids, as opposed to non-recombinant plasmids that are also resistant to ampicillin.</p> Signup and view all the answers

<p>It converts the colourless compound X-gal into a blue-coloured compound, allowing for the identification of bacteria containing recombinant plasmids.</p> Signup and view all the answers

<p>To protect the smaller insulin subunit protein from digestive enzymes within E. coli that may destroy it.</p> Signup and view all the answers

<p>To break down the methionine that was added at the start of the insulin gene, allowing for the separation of the insulin subunit from the ß-galactosidase.</p> Signup and view all the answers

<p>The formation of connecting disulphide bonds, which allows for the creation of functional human insulin.</p> Signup and view all the answers

<p>To produce a large amount of the insulin subunit and ß-galactosidase fusion proteins.</p> Signup and view all the answers