This document discusses recombinant DNA technology. It describes how recombinant DNA technology involves combining DNA fragments from different organisms. The basic steps are: 1) isolating a gene of interest, 2) inserting the fragment into a carrier DNA molecule to generate recombinant DNA, 3) transferring the recombinant DNA into E. coli host cells, and 4) selecting host cells carrying the recombinant DNA. Key tools used are restriction enzymes, which cut DNA at specific sites; vectors like plasmids, which are self-replicating DNA molecules that act as carriers; and host cells like E. coli bacteria.
2. Recombinant dna technologyRecombinant dna technology
A technology which involves theA technology which involves the
combining or rejoining of dnacombining or rejoining of dna
fragment from two differentfragment from two different
organismsorganisms
3. Basics steps in rDNA technologyBasics steps in rDNA technology
1.1.isolation of dna fragment containing aisolation of dna fragment containing a
gene of interest.gene of interest.
2.Generation of a recombinant dna2.Generation of a recombinant dna
molecule by insertion of dnamolecule by insertion of dna
fragment into a carrier dna molecule.fragment into a carrier dna molecule.
3.3.transfer of the rDNA into an e.coli hosttransfer of the rDNA into an e.coli host
cellcell
4.selection of only those host cells4.selection of only those host cells
carrying the rDNA and allowing them tocarrying the rDNA and allowing them to
multiplymultiply
4. Tools of Recombinant DNA techTools of Recombinant DNA tech
1.restriction enzymes1.restriction enzymes
2.vectors2.vectors
3.host cell3.host cell
6. These are the protein molecules also called as theThese are the protein molecules also called as the
molecular scissor becouse they helpto cut the dna atmolecular scissor becouse they helpto cut the dna at
specific sitesspecific sites
properties:-properties:-
1.A restriction enzyme that1.A restriction enzyme that
selectively recognise a specific DNAselectively recognise a specific DNA
sequence and digest any DNAsequence and digest any DNA
fragment containing that sequence.fragment containing that sequence.
2.A modification enzyme that2.A modification enzyme that
adds a methyle grup to one or twoadds a methyle grup to one or two
bases with in the sequencebases with in the sequence
recognised by the enzyme.recognised by the enzyme.
9. plasmidsplasmids
Plasmids are extracromosomal,self-Plasmids are extracromosomal,self-
replicate,usually circular,doublereplicate,usually circular,double
stranded DNA molecule foundstranded DNA molecule found
naturally in many bacteria and innaturally in many bacteria and in
some yeasts.some yeasts.