Genetic Engineering Flashcards
What are the 4 necessities to make recombinant DNA
-restriction Endonucleases (scissors)
-DNA ligase (glue gun)
-Plasmids (workbench)
-E.coli (factory)
What are Endonucleases
Enzymes the bacteria produces to cut up viral genome (bacteriophage)
What is EcoRI
Restriction Endonuclease enzyme and recognise specific DNA sequences and cut them
Characteristics of plasmid
-Circular and double stranded
1. Consist of unique cloning site (EcoRI)
2. Drug resistance gene
3. Origin of replication
What does the drug resistance gene in plasmid allow for
Allows for propagation of plasmid
Where is genome engineering performed in transgenic animals
Performed in Zygote - changes at this stage will be inherited by all cells of the organism (lineage)
What is the ubiquitous promoter (transcriptional)
Drives expression of downstream gene in all tissues at all times - contains a TATA box
What is the polyadenylation signal
Designates site of transcriptional termination - initiates formation of poly (A) tail
-contains two AATAA signals
What are disadvantages of DNA microinjections into zygote
-random transgene integration into genome
-possibility of multiple copies of the transgene
-could disrupt an endogenous gene
What are advantages of genetically engineered embryonic stem cells
-precise integration into genome
-single copy integration
-ideal method to “knock out” genes
What are embryonic stem cells (ES)
-Immortal cell cultures derived from blastocyst embryos
-unlimited growth potential
-can be genetically manipulated
What is the TATA box
Found in the promoter region of the gene and involved in initiating gene transcription
What is a result of gene targeting with CRISPR/Cas9
Increased speed and efficiency of precise genome engineering
What does T4 DNA ligase do
Catalyses formation of phosphodiester bonds in double stranded DNA molecules
-requires 5’ phosphate and 3’ hydroxyl group to do catalysation
-requires ATP as co-factor
