Microbiology Chapter 12 - Modern Application

Biotechnology

The science of utilizing living systems to benefit human kind

Genetic Engineering

The ability to alter an organism’s genome

Recombinant DNA Technology

Technology that allows for advances in genetic engineering by enabling recombinant DNA molecules with new combinations of DNA

Molecular Cloning Methods include…

Restriction Enzymes (to cut foreign DNA and plasmid vectors)

Plasmid Vectors

non-viral DNA molecules that contain no proteins

Restriction Enzymes

Enzymes produced as protection mechanism | cuts and destroys foreign cytoplasmic DNA — commonly a result of a bacteriophage infection | Will cut DNA at recognition site that’s usually Palindromic

Palindromic

DNA sequence typically between 4-6 base pairs in length | Palindrome - sequence of letters read the same forwards and backwards

Plasmids

Small pieces of (typically) circular, double-stranded DNA that replicate independently of the bacterial chromosome

Vectors

DNA molecules that carry DNA fragments from one organism to another

Genomic Libraries

Can be made by cloning genomic fragments from one organism into plasmid vectors or into bacteriophage

cDNA Libraries

Can be made to represent the mRNA molecules expressed in a cell. Lab mRNA is used as a template to reverse transcriptase and make complementary DNA.

Benefit of cDNA

Contains DNA from solely the expressed genes in the cell (due to introns being cut out from cDNA)

Transfection can be achieved by…

Electroporation, Microinjection, Gene guns, Shuttle Vectors, and Viral Vectors

Viral Vectors

Viruses like Adenoviruses are used to deliver foreign DNA to targeted cells

Restriction Fragment Length Polymorphism (RFLP)

Allows for the visualization by agarose gel electrophoresis of distinct variants of a DNA sequence caused by differences in restriction sites

Polymerase Chain Reaction

Allows for rapid amplification of a specific DNA sequence | Occurs over multiple cycles | THREE STEPS: Denaturation, Annealing, and Extension DAE

Science of Genomics

Allows researchers to study organisms on a holistic level | Has great potential for future medical advancements through the study of the human genome

Gene Therapy

So far, have been proven to be relatively inaffective, with exceptions in: cystic fibrosis and adenosine deaminase (a type of SCID)