Recombinant DNA And Cloning Vectors Flashcards
What is a vector?
A type of recombinant tool that is used to transfer DNA into a biological system
What are the types of recombinant vectors?
Plasmids, bacteriophages, viruses and artificial chromosomes
What makes plasmids good recombinant vectors?
They’re transferable by different means including transformation and conjugation
Where are plasmids found?
In most bacteria (not all)
How do bacteriophages transfer the information?
Transduction
What are non-primate lenti viruses?
Vectors used to integrate DNA into mammalian cells
What are baculoviruses?
Vectors that are used in combination with recombinant expression in insect cells
What are features of plasmids?
They can be linearised at one or more sites in non-essential stretches of DNA
Can have DNA inserted into them
Can be recircularised without the loss of ability to replicate
Can replicate lots within a host cell
How do you confirm the success of recombination?
Restriction mapping the clone, then isolating the colonies, culturing and then purifying it
Why are plasmids used in genomics?
To add or modify control elements, express a recombinant gene, or alter the properties of the gene product
Why would you need to alter the properties of a gene product?
To make it secrete extra-cellularly or into the periplasmic space,
To fuse it to a peptide tag or other protein
to make it useful as a therapeutic
What are the control elements needed for expression in bacteria?
Gene coding sequence, shine-dalgarno sequence, bacterial promoter and a transcriptional terminator
What is a constitutive promoter?
A thing that allows a culture of cells to express the foreign protein to a high level
Which type of promoter is always on?
Constitutive
What do inducible promoters do?
Allow large cultures of a thing to be grown without expressing the foreign protein
