Recombinant DNA Technology Flashcards
Who won the Nobel Prize for the first recombinant DNA experiment?
Paul Berg
(1980)
What is Transformation?
Genetic alteration of a cell resulting from the direct uptake and incorporation of exogenous genetic material from its surroundings
How many types of Restriction Enzyme are there?
3
Which type of restriction enzymes are most often used in genetic engineering?
Type II
What modification protects DNA sites from restriction endonuclease?
Methylation
Who made the observation that phage grown in one bacterial host fail to grow in a different “restrictive” bacterial host?
Salvador Luria
Who defined the molecular basis of restriction and modification?
Werner Arber
(1962)
Who characterised the first restriction endonuclease?
Matt Meselson & Bob Yuan
What DNA ligase is used in recombinant DNA?
T4 DNA Ligase
What are the essential features of a plasmid vector?
- Origin of Replication
- Antibiotic Selection
- Multiple Cloning Sites
Who first indentified EcoRI?
Herbert Boyer
How do Restriction Enzymes work?
Cut dsDNA at specific sequence creating blunt or sticky ends
What was the first recombinant DNA experiment?
Inserting new genetic information into DNA of simian virus 40 to produce circular SV40 DNA
What genes does circular SV40 DNA contain?
A molecule containing lambda phage genes and galactose operon of E. coli
What was the first cloning experiment undertaken?
Cloning of frogs
What was organised in 1975 due to fear of recombinant DNA technology?
A landmark conference in Asimolar California
What are the THREE tools of molecular biology?
- Transformation
- Restriction enzymes & Ligase
- Plasmids
Is DNA parallel or anti-parallel?
Anti-parallel
What are bacterial cells that can take up DNA known as?
Competent cells
What are some bacteria that are naturally competent?
Pneumococcus & Neisseria
How can E. coli become competent?
CaCl2 treatment and electroportation
How many enzymes are commercially available?
More than 240
How are restriction enzymes named?
Named for the organism they were discovered
What are pallindromic sequences?
DNA sequence that is the same forward and backwards but on opposite strands
What type of DNA fragment ends are more commonly used in genetic engineering and why?
Sticky ends because of their complimentary tails
What is the size range of plasmids and do they carry essential genes?
1kb to 1.5Mb
They don’t carry essential genes, they carry virulence factors like antibiotic resistance genes
What was the first plasmid vector?
pSC101
(SC = Stanley Cohen)
What was the first biotechnology company?
Genentech
Founded by Herbert Boyer
What was the first recombinant DNA experiment using restriction enzymes?
Newly constructed plasmid inserted into E. coli by transformation
In what plasmid were synthetic genes for human insulin cloned?
In plasmid pBR322
What are genomics?
Identify gene variants in populations
What are the TWO enzymes needed for gene cloning?
- Restriction enzymes
- DNA ligase
