Chapter 3 and 4 Flashcards
endonuclease
enzyme that breaks the phosphodiester bond between two nucleotides in a polynucleotide chain
Recognition site
specific target sequence of DNA upon which restriction endonucleases act
Restriction endonuclease:
any enzyme that acts like molecular scissors to cut nucleic acid strands at specific recognition sites.Also known as restriction enzyme
Ligase
enzyme that joins molecules, including DNA or RNA, together by catalysing the formation of phosphodiester bonds
Polymerase:
enzyme that synthesis a polymer from monomers such as forming a DNA strand from nucleic acids
Primer
short single strand of nucleic acids that act as a starting point for polymerase enzymes to attach
CRISPR-Cas9:
a complex formed between gRNA and Cas9 which can cut a target sequence of DNA. Bacteria use this complex for protection from viruses and scientists have modified it to edit genomes.
Cas (9)
Endonuclease that creates a blunt end cut at a site specified by gRNA
CRISPR
”clustered regularly interspaced short palindromic repeats”,found in prokaryotes that protect them against viral invasion
Spacer
short sequences of DNA obtained from invading bacteriophages that are added into the CRISPR sequence
Genetic modification
manipulation of an organisms genetic material using biotechnology
crispr RNA (crRNA)
made up of spacer and repeat which are transcribed and cleaved to produce the “mugshot” for cas9 to cut DNA
TracerRNA
consists of complementary sequence to the crna repeat which enables the 2 molecules to bond, TracerRNA is responsible for binding tightly with Cas9 to establish the CRISPR-Cas9 complex
Genetic modification:
manipulation of an organisms genetic material using biotechnology
Deleterious mutation
A change in DNA that negatively affects an individual
Gene therapy
repairing genetic mutations by replacing a defective gene with a healthy one
Polymerase chain reaction (PCR)
laboratory technique used to produce many identical copies of DNA from a small initial sample
Taq polymerase
heat resistant DNA polymerase enzyme.
Gel electrophoresis
Technique used to separate DNA fragments based on their molecular size
Plasmid
a small circular loop of DNA separate from the chromosome, typically found in bacteria
Recombinant plasmid
:a circular DNA vector that is ligated to incorporate a gene of interest
Bacterial transformation:
the process by which bacteria take up foreign DNA from their environment
Genetic modification
manipulation of an organisms genetic material using biotechnology
Gene of interest
a gene scientists want to be expressed in recombinant bacteria
Vector:
a means of introducing foreign DNA into an organism.Plasmids are a popular vector in bacterial transformation.
Plasmid vector
a piece of circular DNA that is modified to be an ideal vector for bacterial transformation experiments
Antibiotic resistance gene
gene that inhabits antibiotic resistance
Origin of replication (ORI)
a sequence found in prokaryotes that signals the start site of DNA replication
Reporter gene
gene with an easily identifiable phenotype that can be used to identify whether a plasmid has taken up the gene of interest
Heat shock
:method that involves rapidly increasing then decreasing the temperature to increase the permeability of plasma membrane to enhance the likelihood of bacterial transformation
Electroporation
a method that involves delivering an electric shock to bacterial membranes to increase their membrane permeability
Genetic engineering
:the process of using biotechnology to alter the genome of an organism
Gentically modified organism (GMO)
organism that has genetic material that has been altered using genetic engineering technology
Host organism
organism which researchers wish to genetically modify
Cisgenic organisms
Gentically modified organism that cotains foreign gentic material from a sexually compatable donor organism, typically from the same species
Transgenic organism:
Genetically modified organism that contains foreign genetic material from a separate species.
