after gene isolatin

give example of restriction enzyme

EcoRI

what is a palindromic repeat and why is it useful?

characteristic of recombinant DNA

allows recognition enzymes to recognise and bind to specific sites on DNA

what does ‘sticky’ mean

restriction enzymes cut target DNA fragment + plasmid at specific locations

creates sticky end that binds two fragments

what does a plasmid do in recombinant DNA technology

serves a vector

ligation

insertion of gene into plasmid using DNA ligase

what is transformation

introducing plasmid with gene insert into host organism

e.g. E coli bacteria through heat shock

what is an open reading frame

sequence of RNA or DNA that can be translated into a protein

begins w start codon, ends with stop codon, no interruptions

give 3 applications for GFP

• reporter gene

• tracker for medicinal purposes

• protein localisation in live cells

what is a plasmid

short circular dna in bacteria

what is heat shock

example of transformation

use heat to introduce plasmid w gene insert of E coli bacteria to host organism

explain how you can identify which E.coli colony (after transformation + overnight incubation) contains the target plasmid with the inserted gene. describe the process

• antibiotic resistance screening

• blue-white screening

• colony pcr

• dna sequencing

explain why introns need to be removed before ligation

cant splice its RNA, results in incomplete or nonfunctional protein

how can you remove the introns from the gene isolated from Aequorea Victoria?