DNA sequence produced artificially by joining pieces of DNA from different organisms
Genetic engineering
Technique of altering the genetic makeup of cells or organisms by deliberately inserting, removing, or altering individual genes
GMO
Microorganism, plant, or animal whose genetic material contains a piece of foreign DNA, created using genetic engineering
Creating recombinant DNA
1. DNA fragments containing the gene of interest (transgene)
2. Plasmid DNA
3. Restriction enzyme
4. DNA ligase
Recombinant DNA technology
1. DNA cleavage or cutting by restriction enzymes
2. Choosing a suitable vector or vehicle
3. Linking the gene of interest with the vector
4. Recombinant plasmid transfer into a host cell
5. Method of selection to test which cells contain the gene of interest
6. Sequencing of the gene to find out the protein's primary structure
Gene cloning
Process of synthesizing multiple copies of a particular DNA sequence using a bacteria cell (i.e. plasmid) or another organism as a host
Transformation
Introduction of foreign DNA into cells
Cloning
Transformed bacterial cell is grown in culture to multiply, the recombinant plasmid in each cell also multiplies
Methods to introduce plasmids into host organisms
Gene gun
Heat shock transformation
Methods to screen recombinant cells
Selection of plasmid DNA containing cells
Selection of transformed cells with the desired gene
PCR detection of plasmid DNA
The ability to directly modify the genome of an organism through genetic modification has been made possible in recent years due to developments in recombinant DNA technology
Molecular cloning requires techniques used in host species to create recombinant DNA and promote its replication
