Recombinant DNA technology Flashcards
What is the central dogma?
The flow of DNA to protein, via mRNA
What is replication?
using DNA polymerase to make identical DNA molecules
What is transcription?
using RNA polymerase to make RNA is using DNA as a template.
What is translation?
where proteins are made using mRNA
What is the start codon?
Methionine ATG
What is a gene?
consists of regulatory domains and an open reading frame that contains exons and introns.
What are regulatory domains?
Promoter - tells the RNA polymerase excatly where to start trasncribing
Activator
Silencer
Terminator - where transcription stops
How do we know the genes are eukaryotic?
Eukaryotic genes have exons and introns
Prokaryotes only have introns
What is the difference between a start codon and a promoter?
start codon comes directly after the promoter.
Start codon - initiates translation
Promoter - initiates transcription
How can we manipulate DNA?
Using restriction enzymes, DNA ligase and plasmids.
Bacterial restriction enzymes cut DNA molecules at specific DNA sequences called restriction sites = makes many cuts resulting in restriction fragments
Most useful enzymes cut the molecule in a staggered way producing sticky ends that bind to the complementary sticky ends of other fragments.
DNA ligase seals the bonds between fragments.
What is plasmid?
Small circular piece of DNA that can replicate independently of the chromosome.
What is an antibiotic resistance marker?
Identify organisms that contain this plasmid
What is the origin of replication?
DNA sequence where plasmid replication starts.
What is the MCS?
Mulitple cloning site
a short DNA sequence containing many unique restriction enzyme-cutting sites for different options for cloning the DNA of interest into the plasmid.
WHat is an insert?
Pieces of DNA that have ligated into the plasmid, in the MCS
What is the promoter region?
Drives transcription of insert DNA
Leads to the production of the recombinant DNA protein.
Also can lead to constitutive expression, expression at certain growth phases or be induced by chemicals.