How can plasmid be used to transfer an isolated human gene into a bacterium?
Don’t worry its not to hard i did this in my A-levels, first you need to isolate the human gene this can be done via two ways 1) use restriction endonuclease to cut the gene from a chromosome. (produces ‘sticky ends’) 2) use reverse transcriptase to make a copy of the required gene after the gene is isolated needs to be inserted into a suitable vector which is the plasmid (plasmids occur naturally in bacteria).The plasmid should be cut with restriction endonucleases to ‘open’ it up as it is a circular double standed DNA molecule. An enzyme called DNA ligase helps bond the isolated gene to the plasmid. Plasmid with the target gene is now decribe as recombinant. The plasmid is then put in culture with the bacteria and they enter the host. ( by using Ca ions or an electric shock to disrupt the cell membrane of the host bacteria.) Hope that helps a bit.
