Genetic Engineering Flashcards
Recombinant DNA
Combining 2 or more pieces of DNA not normally found together by artificial means
Restriction Enzymes
- Bacterial defense mechanism -> recognize short sequences of double-stranded DNA as targets
- All have specific consensus sequence -> cleaving to a different base
- 2 Different enzymes can give rise to compatible sticky ends
Gel electrophoresis
- DNA restrictive fragments placed ina well of an agarose gel -> charged
- Molecules move through pores
- Smaller travel further
Southern blot
- Target and probe combined by heat/ alkali then reannealed
- Transfer to nitrocellulose
- Hybridize with labeled DNA/RNA probe
- This gives an audiodiagram
Limitation of plasmids as vectors
Plasmids are limited in amount of DNA they carry -> yeast artificial chromosome 1000 kb
Types of cell transfection
Transient and stable
Difference between types of cell transfection
- Transient -> cells only keep inserted genes for a couple of generations
- Stable -> genes integrated into genome -> constant production of protein
Xenotransplantation
Transfer of living cells from one organism to another
CRISPRs
Clustered regularly interspaced short palindromic repeats
- Bacterial and Archean defence mechanism against plasmids and phages
Cas9
CRISPR associated protein 9, a nuclease
PAM sequence
DNA sequence immediately following the sequence targeted by Cas9
Ethical considerations of GMO’s
- Risk assessment and control measures
- ST + LT impact
- Balanced and informed decision making processes
Western Blotting
- Hybridize RNA polynucleotide
- Mixture of proteins -> separate on gel and transfer onto membrane
- Incubate protein sample with antibody 1 probe
- Ligand-bind with antibody
- If visualized then can view if antibody has bounded to antigen
- Quantify protein -> monomeric, dimeric
Proteomics
Study of protein population: analysis of genome compliments of proteins from DNA -> RNA -> protein