7.3.2 - Transformation and vectors Flashcards
What is a transformed cell?
Host cells that take up the vector.
Describe the process of getting a plasmid into a cell.
Place cells in ice cold calcium chloride solution to make cells more permeable.
Plasmids are added to the mixture and is heat shocked (42 degrees for 2 mins) which encourages the cells to take up the plasmids.
What are 3 reasons why a host cell will not tale up the plasmid.
The recombinant plasmid can’t enter the cell.
The plasmid re-joins before the DNA fragment has entered.
The DNA fragment sticks to itself, rather than inserting into the plasmid.
What can marker genes be used for?
To identify which bacteria successfully took up the recombinant plasmid.
What are 3 common marker genes?
Antibiotic resistance genes
Genes coding for fluorescent proteins.
Genes coding for enzymes.
Describe the process of using the fluorescent protein gene in identifying a transformed cell.
Marker gene inserted into the vectors at the same time as the DNA fragment. DNA fragment in the middle of fluorescent gene. Host cells grown on agar and each cell divides. The transformed cells will not be fluorescent as the gene has been interrupted by the DNA fragment.
Describe the use of the gene machine.
Scientists examine amino acid sequencing in the primary structure of a protein and then work backwards to work out the mRNA sequence and therefore the DNA sequence.
Give an example of in vivo cloning and in vitro cloning.
In vivo - gene markers and plasmids
In vitro - PCR
Why is it useful to silence some genes?
Investigating diseases and testing potential treatments.
Preventing a disease from further replicating.-
