9. Applications Of Microbial Genetics

Crizelda D. Liwanag, MSc, RMT

Genetic Manipulations Applications in Diagnosis

Applications in Diagnosis 1. Detection of DNA • •

• • •

Electrophoresis Southern Blotting

PCR DNA Fingerprinting DNA Sequencing

•3. 2. 4.

Chromosomes, Each short piece which is used range asseparated a in template size from to50 generate million toa 250 set of million The fragments in a set are byof gel electrophoresis New fluorescent dyes allow separation all four fragments in athat bases, fragments must that first differ be broken in length into from much each shorter other pieces by a single (subcloning base (separation step). single on the step). will be lane identified ingel. a later step (template preparation and sequencing reaction steps).

5.7. Automated 6. The After final thebase bases sequencers at the are end "read," analyze of each the computers resulting fragment are electropherograms, isused identified to (base-calling and assemble the output the step). isshort a four-color This sequences process recreates chromatogram (in blocksthe of original about showing 500 sequence peaks basesthat ofeach, represent As, Ts, called each Cs, the and of read the Gs four for length) each DNA short bases. into piece long continuous generated stretches in the first step. that are analyzed for errors, gene-coding regions, and other characteristics.

Genetic Manipulations Applications in Therapy

Applications in Therapy 1. Mutagenecity and antimutagenecity assays 2. Recombinant DNA technology 3. Gene transfer and movement 4. Gene therapy 5. RNA interference 6. Stem cell and organ cloning 7. GMO’s

Gene therapy

Gene therapy

• Insert a working gene

Gene therapy

• Use virus

Gene therapy

• Like an envelope carrying a letter

Gene therapy

• Gene randomly inserts itself into genome

Gene therapy

• It can now be read (correct instruction)

RNA interference

RNA interference

• Suppressors

RNA interference

• From viruses (replace suppressors)

RNA interference

• New genes cause cancer

RNA interference

• Small interfering RNA binds to mRNA of infected genes

Stem cell therapy

• Stem cells

Stem cell therapy

• Trick stem cells of a different organ into regenerating an organ

Stem cell therapy

• Damaged areas of the heart

Stem cell therapy

• Black dots representing areas injected with stem cells

Stem cell therapy

• Stem cells from femur BM are transferred to the heart and they mimic the cells

Stem cell therapy

• Purple area shows healthy heart

Stem cell therapy

• Frog’s eyes grown from stem cells

Stem cell therapy

• 1st to grow sensory organs from embryonic stem cells

Stem cell therapy

• Frogs that cannot see are lighter in color

Stem cell therapy

• Clone of own embryo (to solve histocompatibility and immunologic tolerance problems)

Stem cell therapy

Embryonic stem cell cloning

• Co-creator of DOLLY

Embryonic stem cell cloning

• Trying to clone a human being

Believes cloning will be done

What we still DON’T know, even with the full human sequence in hand: • Gene number, exact locations, and functions • Gene regulation • DNA sequence organization • Chromosomal structure and organization • Noncoding DNA types, amount, distribution, information content, and functions • Coordination of gene expression, protein synthesis, and post-translational events • Interaction of proteins in complex molecular machines

What we still DON’T know, even with the full human sequence in hand: • Predicted vs experimentally determined gene function • Evolutionary conservation among organisms • Protein conservation (structure and function) • Proteomes (total protein content and function) in organisms • Correlation of SNPs (single-base DNA variations among individuals) with health and disease • Disease-susceptibility prediction based on gene sequence variation • Genes involved in complex traits and multigene diseases • Complex systems biology including microbial consortia useful for environmental restoration • Developmental genetics, genomics