Biotechnology/Genetic engineering Flashcards
Biotechnology
Use of microorganisms for human benefit
Not a new science- early civilization made wine/bread
Genetic Engineering
Manipulation of genes/DNA
New science
Human insulin w/ aid of microbes
Diabetes
Not able to produce insulin
Insulin used to be isolated from cows; found out pig insulin is alot better because more similar to human insulin.
Retroviruses
RNA as genetic material and reverse transcriptase enzyme [make DNA from mRNA]
Restriction enzymes
Digest or cut DNA ; break DNA at specific places allowing for scientists to modify DNA
Mechanism of defense for bacteria; remove genetic material from viruses
Recombinant DNA
From 2 sources combined with use of Ligase
Gel Electrophoresis
Determine size of DNA molecules / fragments using Standards
Fragments loaded into wells in a porous agarose gel - gel floats within chamber between two electrodes
Electric current passed, DNA fragments move toward + charged end
Smaller fragments move faster & further
Why is DNA attracted to positive end of electric current
DNA has - charge because of phosphate groups in sugar phosphate backbone, attracted to + end
DNA sequencing
Determines nucleotide sequence in DNA
1) PCR + Gel Electrophoresis
PCR
Amplifies a lot of DNA from small sample
1) Denaturation - heat @ 95 degrees to separate strands (break H- bonds)
2) Annealing - cooled to 60 degrees - primers bind; process if DNA polymerase starts
3) Extension - 72 degrees Taq Polymerase (heat stable DNA Polymerase) to extend primers and form copy strand on template strand
Thermocycler
Continuous change/recycling of temp in PCR.
Cloning into a plasmid
Small piece of circular DNA found in bacteria
Gene of interest (ex: insulin) incorporated with plasmid - recombinant DNA molecule
Plasmid - bacteria & Gene of Interest - human
Bacteria express human gene
Applications of Recombinant DNA
Gene of interest:
- pest resistance
- alter bacteria for cleaning up of toxic waste
- protein dissolves blood clots in heart attack therapy
- protein used to make snow form at higher temperature
Bacterium - plasmid isolated - gene inserted into plasmid- plasmid inserted into bacterial cell-cloning
Transgenic Plants
Genes from other organisms (recombinant DNA)
transgenic = animal/plants and express genes & new traits from other species
Gene Therapy
Altered virus in lab is mixed w/ patient cells = altering patient cells, inject back into patient, desired protein or hormone produced in patient.
Virus - vector
