BIOTECH GENETIC ENGINEERING

• Biotechnology refers to technology used to manipulate DNA
• The procedures Biotechnology are often referred to as Genetic Engineering
• Breakthroughs of Genetic Engineering:

  • CRISPR/Cas 9 was successfully discovered and is used as an editing tool to modify genomes for medicinal purposes
• Genetic engineering made cloning possible, successfully cloning a mammal named "Dolly" from an embryo cell
• Tools used in Genetic Engineering:

  • Polymerase Chain Reaction (PCR):
  • Developed in 1983 by Kary Mullis
  • Multiplies DNA exponentially for 25 to 75 cycles, each taking only a minute
  • Used to amplify a single copy or a few copies of a DNA segment across several orders of magnitude
• Gel Electrophoresis:

  • Used for various purposes, from viewing cut DNA to detecting DNA inserts and knockouts
  • Used to estimate the molecular weight of proteins and nucleic acids, purification of isolated proteins, monitoring changes of protein content in body fluids, and more
• DNA Ligase:

  • Enzymes that can create covalent bonds between nucleotide chains
  • Often utilized in genetic research to link two or more individual strands of DNA, create a recombinant strand, or close a circular strand cut with restriction enzymes
• Polymerases:

  • Groups of enzymes that catalyze nucleic acid molecules' synthesis
  • Three critical polymerases:

  a. DNA-dependent DNA polymerase that replicates DNA from DNA

  b. RNA-dependent DNA polymerase (reverse transcriptase) that transcribes DNA from RNA

  c. DNA-dependent RNA polymerase that transcribes RNA from DNA
• Prokaryotic Host:

  • Able to multiply plasmids (along with foreign DNA) to produce millions of copies. Referred to as a colony or in short, a clone
  • Escherichia coli was the first organism used in DNA technology experiments and continues to be a common host choice
• Eukaryotic Host:

  • Preferred for producing human proteins due to their complex structures
  • The most commonly used eukaryotic organism is the yeast, Saccharomyces cerevisiae
• Selection of Small Self-Replicating DNA:

  • Small circular pieces of DNA capable of self-replication are known as plasmids, often used as vectors to transport genes between microorganisms
  • Viral (bacteriophage) DNA and cosmids (recombinant plasmids containing bacteriophage genes) can also be used as vectors
• Escherichia coli, the simplest Gram-negative bacterium (a common bacterium of human and animal intestines)
• Restriction Enzymes (Molecular Scissor):

  • Enzymes that create one incision on each of the two strands of DNA at specific locations based on the nucleotide sequence
  • Can be separated using gel electrophoresis or chromatography
  • Isolated in 1970 by Hindll
• Gel Electrophoresis is a technique commonly used in laboratories to separate charged molecules like DNA, RNA, and proteins according to their size
• Genetic Engineering is the process by which recombinant DNA technology is utilized to alter the genetic makeup of an organism