#6 help You are a scientist studying the function of a gene you find interesting. You need to produce protein in E.coli so you can purify it and run for functional assay. Which of the following steps is not part of the process of cloning the gene?- Amplifying the target gene through PCR.- Cutting both foreign DNA and plasmid DNA with the same restriction enzyme.- Adding DNA ligase to create a hydrogen bond to seal the nick in the backbone.- Introducing circular recombinant DNA to host bacterial cells.- Selecting the cells with recombinant DNA using selectable markers.

The synthesis of proteins is essential to biological processes and has numerous applications in science, business, and healthcare. Through analysis of the synthesis of particular proteins, provide us insights into the molecular workings behind a range of biological phenomena. Understanding the process of protein synthesis opens up new possibilities for therapeutic interventions by allowing scientists to investigate and create medications that can alter the function of these proteins.The creation of biotechnological products depends on the manufacturing of proteins. The manufacture of medicinal proteins (insulin, antibodies), industrial enzymes, and biofuels is made possible by recombinant DNA technology. Now we will check all the options thoroughly and determine which option is wrong.The target gene is amplified to make many copies using PCR (Polymerase Chain Reaction). A DNA polymerase enzyme, primers that flank the target gene, and nucleotide building blocks are required to do this. A significant quantity of the targeted DNA fragment is produced as the end product. It is an essential process of rDNA technology and the steps are correct.Hence this option is correct.Restriction enzymes also known as restriction endonucleases are used to cut DNA at specific recognition sites. Compatible sticky ends are produced on both the plasmid and the target gene by employing the same restriction enzyme on the vector (target gene) and the foreign DNA (target gene). In order to subsequently connect the foreign DNA with the plasmid, this step is essential. It is an essential process of rDNA technology and the steps are correct.Hence this option is correct.DNA ligase catalyzes the formation of phosphodiester bonds between the 3' hydroxyl end of one nucleotide and the 5' phosphate end of another, effectively joining the two DNA fragments into a continuous, circular molecule.DNA ligase's function is to catalyse the synthesis of phosphodiester bonds—which close the nicks and gaps in between the backbone of DNA strands—between neighbouring nucleotides. It does not, however, produce hydrogen bonds. During the cloning process, the foreign DNA (insert) must be joined to the plasmid DNA (vector) by sealing nicks with DNA ligase. Hence this option is wrong.Using a technique called transformation, the circular recombinant DNA—which now includes the foreign gene of interest within the plasmid vector—is inserted into the host bacterial cells, which are often E. coli. The host cells take up the recombinant DNA and start replicating it along with their own genomic DNA. It is an essential process of rDNA technology and the steps are correct.Hence this option is correct.Selectable markers are frequently present in the plasmid vector, such as genes for antibiotic resistance. Following the introduction of recombinant DNA into the host cells, the cells are cultured in an antibiotic-containing media. Only those cells that have taken up the recombinant DNA (and, therefore, the antibiotic resistance gene) are going to survive. This allows for the selection of cells that have successfully incorporated the foreign gene. It is an essential process of rDNA technology and the steps are correct.Hence this option is correct.Thus the wrong option is "Adding DNA ligase to create a hydrogen bond to seal the nick in the backbone" DNA ligase catalyzes the formation of phosphodiester bond.