Molecular Technologies Flashcards
What are the tools of the trade
DNA libraries
Enzymes used
Vectors
In simple terms what is genetic engineering
Getting piece of dna you want
Getting the piece of DNA into the recipient (bacteria, yeast and other organisms)
Genetic engineering can be achieved in one of 3 ways, what are these ways
- Crosses and selection
- Introduce random mutations and select
- Working with library DNA
What are the stages in genetic engineering
Library
Pcr and add restrictions site
Insert into plasmid
Transform the cell
Select clone
Test clone
Proteins
What does the library stage consist of or mean
Collection of dna/genes from organism of interest
Trying to put the gene we want and put into wanted organsism
Getting the dna libraries can come from 2 sources, what are the 2 types of libraries
Genomic dna - genomic libraries derive from genome contain introns and exons if derived from eukaryotes
Complementary dna (cdna) - derived from mrna
Which library do we want to use for bacteria and why
Cdna library- because you do not want introns and exons
Making the library and getting the right piece depends of what 3 things
Screening
Pcr - to identify bacteria
Luck!
Problem - getting the dna into the cell - how do we cross the membrane
Typically put into bacterium or eukaryotic cell
Packaging the dna - to get into the cell.
Why does dna need to be packaged
So we can get it across the membrane - we use vectors
Vectors - could package DNA into vectors
Give examples of vectors
Plasmids and lambda phage
Vectors - give examples of plasmids
BAC - bacterial aartificial chromosomes
YAC - yeast artificial chromosomes
Fosmid
Cosmid
Which type of plasmid is used when the insert is way too big for plasmid
BAC
Which type of plasmid is used for bigger inserts
YAC
Which type of plasmid is used DNA from phages
Fosmid
