Biotechnology Flashcards
what is biotechnology?
use of biological systems to generate useful products
examples of areas where biotechnology used / can be used?
- transgenic animals
- gene editing
- multigene cassettes
- tissue regeneration
- protein production + protein engineering
- gene therapy
what is exogenous gene expression?
expression of non native genes in organism
expression via plasmids/integration into genome
give examples of expression vectors
- F1 phage origin: viral packaging
- AMP resistance: antibiotic resistance
- ori : origin/replicaion
when cloning any gene, what do you need?
restriction sites at ends of gene
cloning process
- cut both dna mol with same RE
- gel purify 2 fragments - gel electrophoresis
- join fragments using dna ligase
- verify cloning via gel electrophoresis
why do you usually use different RE?
to stop re-ligation + to preserve orientation of insert
gel electrophoresis
used to verify sizes and separate/purify fragments
purified product used for transforming cell
product can = digested with REs to verify that ligation has worked
what do eukaryotes use for strong expression?
viral promoters (CMV/SV40)
what are more specific promoters used for?
lower expression levels
for most biotech processes, what is used?
e.coli
mammalian cell expression system
- correctly folded + fully post-transcriptionally modified
- transported to culture media
available for purification
bacterial/yeast cell expression host
- intracellular expression
- soluble/insoluble
- cell breakage needed
available for purification and modification if needed
give an example of a recombinant vector?
lac operon
what is the lac operon used for?
control elements for heterologous protein expression
