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Medical Biotechnology, step 1.9, Copies of recombinant plasmid cloned by…

- - - - Bacterial cell
      - mRNA extracted from human pancreatic cells
    - - Protein hormones produces in the pancreas.
    - - Become treatable
  - - - PROBLEM IN TREATMENT
        
        Insufficient supply as the used growth hormone extracted from pituitary glands of dead humans
        
        Creutzfeldt-Jakob disease: extraction from pituitary glands were sometimes contaminated with an infectious protein
    - - DNA code (gene) added to bacterium is complementary DNA (cDNA) from mRNA using reverse transcriptase
        
        Before added into vector, another piece of DNA added into cDNA from bacterium E.coli. The piece is the code for a signal sequence (translated becomes a sequence of amino acids)
        
        Hormone is then secreted from bacterium into surrounding medium, making purification a lot easier
        
        Signal sequence is removed by a bacterial enzyme after its release (leaving pure hormone)
  - - - The use of recombinant DNA technology or genetic engineering to overcome genetics disease
    - - germ-line gene therapy
        
        sperms or egg cells are treated
      - somatic gene therapy
        
        only the body cells are altered
    - - Bacterial cell
        
        Bacterial plasmid isolated
        
        Bacterial plasmid cut with restriction endonuclease
        
        DNA for normal CFTP
      - Cells with allele for normal CTFP
        
        DNA coding strand for normal CTFP isolated
        
        Normal CTFP allele inserted into bacterial plasmid using DNA ligase
    - - cystic fibrous transmembrane protein (CFTP)
        
        codes for production of cystic fibrosis transmembrane protein
        
        smooth epithelia and watery mucus
        
        trapping dirt and microorganism
        
        prevent lung infections
      - altered base sequences in gene coding
        
        no protein or faulty protein
        
        too thick and sticky mucus
        
        airways clogged and becomes lung infection
      - lack of CFTP
        
        formation of fibrous cyst in pancreas
        
        interferes with digestion of food
    - - Ex-vivo
        
        outside the organism's body
        
        exp: to treat Severe Combined Immunodeficiency Disease (SCID)
      - In-vivo
        
        Insert RNA of normal allele into retrovirus.
        
        Let retrovirus infect bone marrow cells that have removed from the patient and culture.
        
        Viral DNA carrying the normal allele is inserted into the chromosome.
        
        Inject the engineered cells into the patient.
        
        inside organism's body
        
        exp: to treat cystic fibrosis by liposomes or viral delivery system
  - - - Interferes with the transmission of signals from nerves to muscles
      - Used as a painkiller and provide pain relief