You cannot insert a DNA sequence into a traditional vector if it is over 20 kilobases (kb) in length. The only concern with making a plasmid is that there is a DNA sequence limitation. The target gene here is our SUPER APE gene.
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DNA ligase is an enzyme that takes a free phosphate from the 5' end of one piece of DNA and attaches it to the free –OH at the 3' end of the other piece of DNA.Ībove is a schematic of how most scientists make recombinant DNA plasmids. Instead, we use a "molecular glue" called DNA ligase. Elmer's does not make bottles that small. How do you join the two sequences? Naturally, you join them in the same way you join everything else: glue! Once you have cut your vector in the MCS, or elsewhere in the vector that has a unique restriction site, you need to join it together. could learn something from restriction enzymes. Sticky ends are generally easier to handle, and different restriction enzymes often leave the same overhanging sequences, so you can cut your vector with one enzyme and your DNA insert with another enzyme, and the two sticky ends are then called compatible. Restriction enzymes cut in a way that either leaves sticky ends, where each strand is cut at a different position, leaving 2–5 bases of single-stranded DNA, or blunt ends, where the same spot on both strands is cleaved. MCS sequences have sites for common restriction enzymes, and these enzymes were designed to cut the DNA only in the MCS and nowhere else in the vector. There are thousands of different restriction enzymes that each cleave a specific sequence. Restriction enzymes cleave DNA at specific palindromic sequences, such as GGATCC, GAATTC, and GTTAAC. This is because the target gene will disrupt the lacZ or ccdB gene, and one can easily test the activity of either of these genes by looking for colonies on a plate that lack the relevant gene activity.Ī multiple cloning site is a DNA sequence that contains many restriction enzyme sites. Putting the MCS into a lacZ or ccdB gene allows selection for bacteria that have the target gene inserted into the vector.
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So you want to make an army of super apes, but where do you start? While the following section is completely hypothetical, it shows how scientists make genetically recombinant organisms.ĭisclaimer: do not tell your friends after reading this that you know how to make an army of super apes. And just for fun, let's be evil about it. For now, let's pretend we're scientists making recombinant DNA. We will discuss the advantages and disadvantages of recombinant DNA technology in the "Biotechnology" section. Hey, we at Shmoop admit that super apes are super scary. However, this technology has far more advantages than disadvantages, and the chances of creating super apes that will wipe out humanity are very small. Movies like Rise of the Planet of the Apes, The Island of Doctor Moreau, and Jurassic Park all feature evil scientists doing evil science with evil recombinant DNA. Non- Spider-Man films play off those fears with the evil scientist character. Many people fear that manipulating recombinant DNA is akin to "playing God," and some Making recombinant DNA involves taking a gene, usually from an eukaryote, and putting it into a bacterial plasmid. Unfortunately, that method of transferring spider powers is highly unlikely, knowing what we know about DNA. If you are a Spider-Man fan, you will know the original story had Peter Parker bitten by a radioactive spider. Therefore, in this case, Spider-Man exists because recombinant DNA from spider genes entered the human genome of Peter Parker. In the most recent Spider-Man movie, a mutant spider bit Peter Parker and passed its spider genes to him, giving him super strength, vision, and web-slinging action. Recombinant DNA Spider-Man and Other Examples of Recombinant DNA