recombinant technologys Flashcards
Recombinant DNA technologys
Process of joining two sections of DNA together (can be from different species) and inserting it into a host to create proteins
What is the main tool used in recombiant technologys
The vector which is the method of carrying DNA
Used = plasmids
- double stranded DNA seperate from central nucleus + replicate individually
- contain genes to provide functions
- mostly found in bacteria (occasonally euakaryotes)
What are the key components of a vactor
origon of replication = start point for replication
antibiotic resistance gene = keeps anything carrying vector alive when treated
other gene marker = method to see if cell has uptaken gene (flurences such as GFP)
promotor = specific to each host -> different TF bind to diffferent promotors, enables transcription of gene of interest
recognition sites = sites restrictive enzymes recognize and cut
gene of interest = gene which protein you want inserted
What is the importance of the genetic universal code for recombinant technologies + what is this concept
Genetic universal code: Concept that all organisums read DNA in the same 3 letter codon = amino acid pairing system universally
enables any gene to be correctly interprated in any organisum
Lets up transcibe human genes be read in other organisums -> create proteins
What is the complications with converting human gDNA into a prokaryote for a fucntional protein
eukaryotic DNA has intron which cannot be spliced in a prokaryote -> leads to a non-sence protein
solution:
transcribe DNA to pre-mRNA -> splice to mRNA -> use reverse transcriptase to create cDNA (coding DNA) which doesnt have introns and can be accuartly translated in prokaryotes
What is transformation of a vector
The insertion into a host,
initally into a bacteria for rapid growth and replication of DNA,
- replication kills any bacteria without vector due to antibiotics
How is the gene of interest inserted into the vector
Restrictive enzyme is used to cute at recognition site -> leave an overhand on each side
DNA sequence attached to over hand, secured by DNA ligase
- creates phosphodiesters
