Genetic Engineering

New characteristics

Processed gene

GEO → Genetically Engineered Organism

GMO → Genetically Modified Organism

TO→ Transgenic Organism

1st used by: Jack Williamson in his Dragon Island book

STEPS OF GENETIC ENGINEERING

☆Select desired DNA:

  1. From somatic cell

☆Select the carrier or vector:

  1. E. coli

  2. Agrobacterium tumefaciens

☆Cutting:

  1. Restriction enzyme used

  2. Cut the desired DNA

☆Join:

  1. Ligase enzyme used

  2. Place the desired DNA in the plasmid DNA of carrier

☆Select host and inject:

  1. E. coli used

  2. Electroporation→electric medium through pores

  3. Micro injection→use micropipette

  4. Biolistics→gunshots

  5. Calcium chloride→heat shock

  6. Liposomes

  7. Use Agrobacterium tumefaciens or TMV directly

  8. Mostly heatshock given

☆Analyze:

  1. Place in Agar media (to make the cell grow and express characteristics) and antibiotics (to make it's immunity strong)

  2. Detect and verify if it’s carrying the desired gene

  3. Genetic probe→ metal detector

  4. PCR method→ polymerase chain reaction

  5. Restriction digestion

☆Multiple:

  1. Place the recombinant DNA in desired cell

  2. Tissue culture used to Multiple

PLASMID

Bacterial DNA outside the chromosome

  1. Circular, Double stranded DNA

  2. Self reproduction ability

  3. Small amount of gene

  4. Weight 10⁶-200×10⁶ Dalton

  5. Reproduction by conjugation

  6. Can be cut by restriction enzyme

  7. Contains special chemical substances like colicin, vibriocin etc.

Plasmid types:

  1. F and F’ plasmid (fertility) → transfer genetic material, produces pili

  2. R plasmid (resistance) → antibiotic properties, R6 for 6 antibiotics

  3. Col plasmid (colicin) → destroys E. coli bacteria

RESTRICTION ENZYME

Restricts replication, transpiration

Biological knife/scissors

Cuts specific parts of DNA (restriction site or recognition site)

Also known as restriction endonucleases