In Vivo Cloning Flashcards
True or false: whole organism cloning is the same as DNA cloning
False
What is the basis of In Vivo Cloning
Uses cell to amplify DNA, involves construction of novel DNA molecules by joining DNA from different sources. Product is called Recombinant DNA
Why do we clone DNA (in vivo)
To analyse DNA control sequences Protein, enzyme, RNA func. Investigation Mutation ID Engineering organisms Purify and produce hormones like GH that can’t be synthesised artificially
How is a gene inserted into a plasmid
Plasmids have specifically regions that can be cut. DNA is then inserted into that region and annealed back up
How can we use plasmids (gene insertion) to cure diseases, give an example
For diabetes, Put the gene-inserted (e.g. gene for insulin production) plasmid into bacterium (biological vector) that grows quickly, these will replicate plasmid and express genes inserted into plasmid
Bacteria have restriction enzymes that cut out spec. DNA sequences from plasmids like viral infections or bacteriophages. How do they avoid cutting out their own DNA
They methylate their DNA with methylase which provides it protection against restriction enzymes
What does DNA Ligase do
Sticks sticky ends together (useful when cloning)
Sticks two DNA molecules together in general
Blunt ends and sticky ends can be joined with the aid of the missing complimentary dNTPs
Name some features of biological vectors
Must contain replicon to allow replication in host cells
Has several marker genes
Unique cleave (restriction) sites
Must contain control elements for expression like ribosome binding sites, promoters and terminators
Name some types of biological vector
Plasmids Phases Dismiss Yeast artificial chromosomes Bacterial artificial chromosomes Transposons
True or false: plasmids can replicate independently of host cell
True
How many Kb pairs can a plasmid hold
10Kb pairs
What ways can plasmid DNA be inserted in biological vector
Transformation Electroportation Protoplasm fusion Micro injection Gene gun
