DNA Cloning: Recombinant DNA Technology

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DNA CloningRecombinant DNA Technology

• Recombinant DNA contains DNA from two or more
  different sources
• A vector
• introduces rDNA into host cell
• Plasmids are common vectors
• Two enzymes to introduce foreign DNA into vector
  DNA
• A restriction enzyme - cleaves DNA, and
• A DNA ligase enzyme - seals DNA into an opening
  created by the restriction enzyme

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DNA CloningPolymerase Chain Reaction (PCR)

• Amplifies a targeted sequence of DNA
• Allows thousands of copies to be made of small
  samples of DNA
• Requires
• DNA polymerase
• A supply of nucleotides for the new,
  complementary strand

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Applications of PCRAnalyzing DNA Segments

• DNA can be subjected to DNA fingerprinting
• Treat DNA segment with restriction enzymes
• A unique collection of different fragments is
  produced
• Gel electrophoresis separates the fragments
  according to their charge/size
• Produces distinctive banding pattern
• Used in paternity suits, rape cases, corpse ID,
  etc.

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Biotechnology ProductsTransgenic Animals

• Transgenic Animals
• Gene Pharming
• Use of transgenic farm animals to produce
  pharmaceuticals
• Genes coding for therapeutic diagnostic
  proteins are incorporated into an animals DNA
• The proteins appear in the animals milk
• Plans are to produce drugs to treat
• Cystic fibrosis
• Cancer
• Blood diseases, etc.

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Human Genome Project

• Genome - All the genetic information of an
  individual (or species)
• Goals of Human Genome Project
• Determine the base pair sequence
• Construct a map showing sequence of genes on
  specific chromosomes
• Other species in final stages

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Genetic Profile

• The complete genotype of an individual
• This is the persons genetic profile
• A way of studying how genes work together to
  control the phenotype
• Analyze the genetic profile of many individuals
• Compare their profiles to their phenotypes

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Proteomics

• The study of the structure, function, and
  interaction of cellular proteins
• At least 25,000 of our genes are translated into
  proteins
• The sum total of these proteins is called the
  human proteome
• Understanding protein function is essential to
  the development of better drugs
• Correlate drug treatment to the particular genome
• Increase efficiency and decrease side effects

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Bioinformatics

• The application of computer technologies to the
  study of the genome
• Genomics and proteomics produce raw data
• These fields depend on computer analysis to find
  significant patterns in the data
• Scientists hope to find relationships between
  genetic profiles and genetic disorders

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Gene Therapy

• The insertion of genetic material into human
  cells for the treatment of a disorder
• Ex Vivo
• Children with Severe Combined Immunodeficiency
• Bone Marrow Stem Cells
• In Vivo
• Cystic Fibrosis
• Nasal / Respiratory Spray