circular, double-stranded plasmid DNA (cloning vector) DNA fragment to be cloned
ID: 48955 • Letter: C
Question
circular, double-stranded plasmid DNA (cloning vector) DNA fragment to be cloned recombinant DNA CLEAVAGE WITH RESTRICTION NUCLEASE COVALENT LINKAGE BY DNA LIGASE 200 nm 200 nm STAGGERED END FILLED IN BY DNA POLYMERASE dNTPs 5 GAATT CC GA ATT CC CTTA A CTTA ATP +ligase ATP GAATT GAATT C CT TAA G CTTAAGG (A) JOINING TWO FRAGMENTS CUT BY THE SAME RESTRICTION NUCLEASE (B) JOINING TWO FRAGMENTS CUT BY DIFFERENT RESTRICTION NUCLEASES Figure 8-28 Molecular Biology of the Cell 6e (O Garland Science 2015)Explanation / Answer
This slide shows the process of how the recombinant DNA is formed with new DNA frnagment which has been recieved from the bacteria
plasmid gets cleaved by restriciton nuclease and that makes a space for bacterial DNA to get attached then with the help of DNA ligase it get attached to the plasmid by covalent bonding . this forms recombinant DNA . now whenever this dna will replicate it will have bacterial properties i.e bacterial DNA is replicating with plasmid
The second figure shows how two fragments join. one from 5 to 3 direction adn other from 3 to 5 direction get attached in presence of ligase because the 5 to 3 DNA was having the required opposite bases . so it gets attached
but in the second case sttagered DNA first form the opposite base and then gets attached to its counterpart in presence of ligase. this ocur in presence of different restriction nuclease .
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