1. Tools and techniques of genetic engineering * Enzymes for dicing, splicing, and reversing nucleic acids. * Analysis of DNA. * Restriction endonucleases- recognize specific sequence of DNA and break phosphate-sugar bond. * Liagase- rejoins phosphate-sugar bonds cuts by endonucleases. * Reverse transcript-makes a DNA copy of RNA.
* Analysis of DNA * Gel electrophoresis- separates DNA fragments based on size. * Nucleic acid hybridization and phrobes- probes based pair with complementary sequence used to detect specific sequences. * DNA sequence- reading the sequence of nucleotides in a stretch of DNA. * Polymerase chain reaction- way to amplify DNA.
2. Methods in Recombinant DNA technology: * Concerned with transferring DNA from one organism to another. * Cloning vectors and hosts. * Construction of a recombinant plasmid (vector can be a movement or a viruses) * Characteristics of Cloning Vectors: * Must be capable of carrying a significant piece of donor DNA. * Must be accepted by the cloning host. * Plasmids- small, well characterized, easy to manipulate and can be transferred into appropriate host cells through transformation. * Bacteria phages- have the natural ability to inject their DNA into bacterial hosts through transduction. 3. Characteristics of Cloning Hosts: * Rapid overturn, fast growth rate. * Can be grown in large quantities using ordinary culture methods. * Nonpathogenic * Genome that is well delineated. * Capable of accepting plasmids or bacteriophage vectors. * Maintain foreign genes through multiple generations * Secrete a high yield of proteins from expressed foreign genes. 4. Biochemical Products of Recombinant DNA