RECOMBINANT DNA TECHNOLOGY
por afshan banu
1. GENE CLONNING
1.1. INSERTION OF FORGEIN DNA INTO ANOTHER ORGANISM
2. ENZYMES REQUIRED
2.1. NUCLEASES
2.2. LIGASES
2.2.1. CUTS NUCLEOTIDES DEGRADE NUCLEIC ACIDS
2.2.1.1. EXONUCLEASES
2.2.1.1.1. ACTS AT DNA
2.2.1.2. ENDONUCLEASES
2.2.1.2.1. REMOVE NUCLEOTIDES WITHIN DNA AND DEGRADES NUCLEIC ACID
2.3. END MODYFYING ENZYME
2.3.1. JOIN OR LIGATES THE ENDS OF DNA
2.4. TOPOISOMERASE
2.4.1. ACTS AT THE ENDS OF DNA
2.4.2. MAINTAINS TOPOLOGY OF THE DNA
2.4.2.1. COILING OF DNA
2.4.2.1.1. POSITIVE COILING -TOPOISOMERASE 1
2.4.2.1.2. NEGATIVE COILING - TOPOISOMERSE 2 (DNA GYRASE)
2.5. POLYMERASES
2.5.1. ADDING NUCLEOTIDES TO THE EXISTING PRIMER
3. STEPS INVOLVED
3.1. 1.ISOLATION OF GENE OF INTEREST
3.2. 2.CHOOSING CLONING VECTOR (PLASMID)
3.3. 3.CUTTING OF DNA BY RESTRICTION ENZYME
3.3.1. 3.5 . JOINING OF DNA BY DNA LIGASE
3.4. TRANSFERATION INTO HOST
3.5. SELECTION AND SCREENING OF RECOMBINANTS
4. DNA IS PRODUCED IN VITRO
5. INTO WHICH THE RECOMBINANT DNA IS INTRODUCED
6. HOST ORGANISM
7. DEFINITION
7.1. CUTTING OF DNA AND PASTING TO OBTAIN DESIRED CELL OF INTEREST
7.2. TECHNIQUES INVOLED IN THE CONSTRUCTION,STUDY AND USE OF RECOMBINANT DNA MOLECULES
8. TOOLS NEEDED
8.1. RESTRICTION ENZYMES
8.1.1. CUTS DNA AT SPECIFIC SITES
8.2. CLONNING VECTOR
8.2.1. HELPS IN CARRYING AND INTERGRATINGTHE DESIRED GENE
8.3. DNA LIGASE
8.3.1. JOINS THE ENDS OF DNA