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Study of Cloning Vectors, Restriction Endonucleases and DNA Ligase SlideShare

BPharmacy, 6th Semester, 2021 - Notes

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Study of Cloning Vectors, Restriction Endonucleases and DNA Ligase SlideShare BPharmacy 6th semester 2021 Previous Year's Question Papers/Notes Download - HK Technical PGIMS

1. Study of Cloning Vectors, Restriction Endonucleases and DNA ligase Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 2. What is cloning Features of cloning vector Importance of cloning vector Types of cloning vector Restriction Enzymes Steps involved RE Classes Introduction Types of DNA ligase Mechanism Applications Let’s Learn with Easiest Way Content 2 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 3. Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS What is Vector? PRIOR CONCEPT 3 4. What is Cloning? PRIOR CONCEPT Clothing in biotechnology refers to the process of creating clones of organisms or copies of cells or DNA fragments. Cloning Vector 4 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 5. Cloning Vector PRIOR CONCEPT 5 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 6. Cloning Vector TYPES 6 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 7. Plasmids TYPES - CLONING VECTOR 7 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 8. Plasmids TYPES - CLONING VECTOR Construction of a Plasmid Vector [HD Animation] 8 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 9. Plasmids TYPES - CLONING VECTOR 9 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 10. Plasmids TYPES - CLONING VECTOR 10 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 11. Plasmids TYPES - CLONING VECTOR 11 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 12. Plasmids TYPES - CLONING VECTOR 12 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 13. Bacteriophages TYPES - CLONING VECTOR 13 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 14. Bacteriophages TYPES - CLONING VECTOR 14 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 15. Cosmids TYPES - CLONING VECTOR 15 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 16. Bacterial Artificial Chromosomes (BACs) TYPES - CLONING VECTOR 16 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 17. TYPES - CLONING VECTOR Yeast Artificial Chromosomes (YACs) 17 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 18. TYPES - CLONING VECTOR Retroviral Vectors 18 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 19. PRIOR CONCEPT What are restriction enzymes?  Molecular scissors that cut double stranded DNA molecules at specific points.  Found naturally in a wide variety of prokaryotes  An important tool for manipulating DNA  Most bacteria use Restriction Enzymes as a defence against bacteriophages.  Restriction enzymes prevent the replication of the phage by cleaving its DNA at specific sites.  The host DNA is protected by Methylases which add methyl groups to adenine or cytosine bases within the recognition site thereby modifying the site and protecting the DNA. Biological Role 19 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 20. PRIOR CONCEPT History of Restriction Enzyme  First restriction enzyme was isoltaed in 1970 by Hindll.  He also done the subsequent discovery and characterization of numerous restriction endonucleases.  From then Over 3000 restriction enzymes have been studied in detail, and more than 600 of these are available commercially and are routinely used for DNA modification and manipulation in laboratories. 20 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 21. RE Mechanism of Action Restriction Endonuclease scan the length of the DNA binds to the DNA molecule when it recognizes a specific sequence and makes one cut in each of the sugar phosphate backbones of the double helix - by hydrolyzing the phoshphodiester bond. Specifically, the bond between the 3' O atom and the P atom is broken. 21 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 22. RE Nucleophilic Attack at Phosphorus Atom 22 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 23. RE Palindrome Sequences 23 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 24. RE 24 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS Ends of Restriction Fragments 25. RE 25 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS Blunt Ends 26. RE 26 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS Sticky Ends 27. RE 27 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 28. RE 28 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 29. RE 29 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS Nomenclature of Restriction Enzyme 30. Types Restriction Enzymes 30 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 31. Types Restriction Endonucleases 31 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 32. Types of RE 32 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 33. Types of RE 33 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 34. Types of RE 34 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 35. Types of RE 35 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 36. RE Application of Restriction Enzymes 36 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 37. DNA Ligase PRIOR CONCEPT 37 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 38. DNA Ligase M.O.A The mechanism of DNA ligase is to form two covalent phosphodiester bonds between 3' hydroxyl ends of one nucleotide, ("acceptor") with the 5' phosphate end of another ("donor"). ATP is required for the ligase reaction, which proceeds in three steps: 1. adenylation (addition of AMP) of a residue in the active center of the enzyme, pyrophosphate is released. 2. transfer of the AMP to the 5' phosphate so-called donor, formation of a pyrophosphate bond; 3. formation of a phosphodiester bond between the 5' phosphate of the donor and the 3' hydroxyl of the acceptor. 38 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 39. Mechanism DNA LIGASE 39 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 40. DNA Ligase TYPES 40 1. Bacteriophage T4 DNA Ligase 2. E.coli DNA Ligase Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 41. Bacteriophage T4 DNA Ligase (ATP) TYPES DNS LIGASE 41 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 42. TYPES DNS LIGASE E.Coli DNA Ligase 42 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 43. TYPES DNS LIGASE T4 RNA Ligase 43 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 44. DNA LIGASE Applications 44 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS 45. 45 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS https://pgims.hktechnical.com Downloaded From 46. 46 Study of Cloning Vectors, Restriction Endonucleases and DNA ligase SDPGIPS Rohtak – HK Technical PGIMS



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