Team:Bilkent UNAM Turkey/Transformation


Insertion of nucleic acids within a cell. If it is done in a deliberate manner and the target cell is eukaryotic, the term transfection is used. May occur in nature without human interference – this was the reason the Griffith experiment succeeded, as Streptococcus pneumoniae is generally receptive to foreign nucleic acids. Other cells, however, have to be coaxed into accepting the desired sequence; this is usually accomplished by opening pores in the cell membrane (i.e. By heating or electrocution), using chemical methods to facilitate nucleic acid transfer (i.e. calcium chloride transfection) or mechanical insertion with sufficiently thin needles. Bacteria and single-called eukaryotes can sustain plasmids for large periods of time and can therefore be transfected with plasmids without worry, but in higher Eukarya, plasmids cannot be retained for over c. 48h. If one desires the sequence to be retained and expressed longer periods of time, additional steps must be taken to ensure that the inserted sequence integrates into the genome (this is called stable transfection.) Since we’re working with C. reinhardtii, we need not worry about stable transfection.
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